Anti-infection

Western blot analysis of histone H3 acetylation in the spleen of untreated and vorinostat-treated hNF-E2 tg mice (n = 4 of each genotype).

Three-dimensional responses of MCF7/IGF-1R cells to TAM (1 μM), E2 and IGF-1. Compared to parental MCF7 cells (a), MCF7/IGF-1R cells (b) in three-dimensional (3D) culture formed bigger acini in response to IGF-1 stimulation and displayed significant TAM resistance when treated with TAM (1 μM) + E2 + IGF-1, which was removable by kinase inhibitors BMS-536924, U0126 and BEZ235 (c). Cells (10,000/well) were seeded in 96-well plates. Acini were formed on 100% Matrigel and cultured for 14 days in starving medium containing 2% Matrigel and 5% charcoal/dextran-stripped fetal bovine serum with the treatments as indicated. Concentrations used: TAM (1 μM), E2 (1 nM) and IGF-1 (100 ng/mL). Confocal image original magnification, × 20. Red, rhodamine phalloidin (actin). Blue, Hoechst blue stain. Results are representative of two individual experiments.

Cells were stimulated with TPA (10 nM) for 15 min in the presence of the indicated concentrations of U0126. Samples were collected and analyzed by Western blot to detect phosphorylated p42/p44 MAPK.

CP-690550 reduced the severity of ischemic damage. (A) CP-690550 (10 μM) suppressed IL-17 production by cdT cells and activated memory T cells. cdT cells and CD4+ CD44+ memory T cells isolated from C5BL6J mice through flow cytometry were stimulated for 24 h with plate-bound monoclonal antibodies to CD3 (2 ng/ml) and CD28 (1 ng/ml) in the presence or absence of IL-23 (25 ng/ml). IL-17A production and IL-17A expression level were measured by quantitative RT-PCR (a) and ELISA (b). (B) CBF reduction after brain ischemia. (C) Time-dependent changes in neurological score. *p < 0.05. (D) Infarct volume as visualized through TTC staining on day 3 in CP-690550 (CP)- and vehicle-treated mice.

 

(C) In vivo treatment of Tg:Pomc-Pttg;Pomc-eGFP embryos with small-molecule CDK inhibitors (50 μM) or 0.2% DMSO as control from 18 to 40 hpf. One hundred to one hundred fifty embryos were treated with each compound. Representative images of live embryos are shown with gross morphology (Right) and pituitary Pomc-GFP-positive cells at higher magnification (Left) at 40 hpf. Embryos exposed to flavopiridol developed early developmental defect before pituitary POMC cell ontogeny occurs. (D) Relative expression of pituitary Pomc-eGFP fluorescence analyzed using Volocity 5.2 software (Improvision; mean ±SE of relative expression, n = 7). (E) R-roscovitine specifically suppresses expansion of pituitary POMC cells overexpressing zPttg from 18 to 48 hpf. Double transgenic Tg:Pomc-Pttg;Prl-RFP embryos were generated by breeding Tg:Pomc-Pttg fish with a previously generated PRL-RFP transgenic line, in which RFP was targeted to pituitary lactotrophs by a zebrafish Prolactin promoter (34). Representative fluorescent microscopy of pituitary POMC-eGFP (a and b) and PRL-RFP (c and d) expression in live Tg:Pomc-Pttg; Pomc-eGFP and Tg:Pomc-Pttg;Prl-RFP embryos treated with 0.2% DMSO (a and c) or 50 μM R-roscovitine (b and d). (F) Relative expression of pituitary POMC-eGFP or PRL-RFP fluorescence were analyzed (mean ±SE of relative expression; n = 10). Results represent one of three similar experiments;*P < 0.02 and **P < 0.000005. (Scale bar, 50 μm.)

CCR5 antagonists block FBS-induced breast cancer cell invasion. 3D reconstruction of FBS-induced invasion into collagen gels by Hs578T (A) or SUM-159 (C) breast cancer cells in presence of CCR5 antagonists (100 nmol/L). The corresponding quantifications (mean ±SEM, n = 3) and analysis (Bonferronit test) are displayed in B and D.

Comparative efficacy of anticancer therapies (Flavopiridol, vincristine, daunorubicin, et al.) in NMC vs non-NMC cell lines. Mean IC50 (± s.e.m.) of the indicated agents in three NMC (PER-403, PER-624, and PER-704) and two non-NMC cell lines (PER-535 and SAOS2), ***P<0.001, unpaired t-test, corrected for multiple testing.

A549 cells were treated with G15 (a specific antagonist of GPR30, 1 uM), AG1478 (a potent antagonist of EGFR, 10 uM), BPA (10^-5 M) alone for 15 min or BPA after a 90-min pretreatment with G15 or AG1478 for 15 min. Then the expression of p-ERK1/2 and total ERK1/2 were measured by western blot analysis.

DNA-PKcs suppression mediated ROS production and GSH content in HepG2 cells exposed to CDDP and 5-Fu. a DNA-PKcs inhibition promoted ROS production in HepG2 cells treated with indicated concentrations of CDDP and 5-Fu. DCFH-DA fluorescent analysis was performed to assess the ROS level. Data presented were mean ?SD of three independent experiments.

(B and C) sh-scrambled or sh-ck2a–transducted L929 cells (B) and Raw cells (C) were stimulated by DMXAA (100 μg/ml) for various times. Cytosolic and nuclear extracts were prepared as described in Materials and Methods. Five percent of the cytosolic proteins and 20% of the nuclear proteins were resolved by 10% SDS-PAGE. Subsequently, immunoblotting was conducted by indicated Abs. The amounts of Tubulin and Lamin B1 in cytosol versus nuclei detected by respective Abs were used as internal control for fractionation.

Effect of triciribine on the migration of (A) FaDu and (B) Hep2 cells. The cells were treated with 5 µM triciribine for different periods of time. *P<0.05 vs. control. Hpf, high-power field.

Sensitivity of AML cells to conventional induction treatment and small-molecule p53 activators. Cell viability in OCI-AML3 cells treated for 24 hours with cytarabin/idarubicin (CI) and Nultin-3A (Nut; A), CI and Leptomycin-B (LMB; B), or Nut and LMB (D), respectively. CI was tested at 0, 100, 200, and 300 nmol/L CI; Nutlin-3A at 0, 2.5, 5, 7.5; and LMB at 0, 2, 8, 32 ng/mL in dosages 0, 1, 2, and 3, respectively. Cell viability in normal and AML bone marrow cells.

Cells were treated with brefeldin A or manumycin A, and the resulting supernatant was collected after 48 h for exosomal preparation (lanes 1 and 2), or exosomes obtained from C81 cells were trypsin-treated or freeze/thawed (F/T) and then trypsin-treated (lanes 3 and 4). Lanes 5 and 6, input exosome controls from C81 or CEM cells, respectively. Resulting exosomes were assayed for the presence of Tax by Western blotting.

HepT1, HepG2, and HuH6 cells were treated with increasing doses of aprepitant or SP [100 nM] and Western blot analysis was performed for the apoptotic markers PARP, Caspase-3, and BID.

Phenotypic effect of Genetic or Pharmacologic Compromise of the 477 Hsp70-StiA-Hsp90 Complex. The impact of each genetic modification on radial growth, conidiation, and response to various stress conditions was assessed after inoculation of a suspension of 104 conidia on glucose minimal medium (GMM) agar plates and incubation at 37ºC for 5 days.

The Nucleotide analog reverse-transcriptase inhibitor Tenofovir disoproxil fumarate was added to TZM-bl cells. Cells were inoculated with 0.05 ng mock-exposed and semen-exposed HIV, and 0.5 ng HIV as infectivity-matched control. Infection rates were measured 3 days post infection by measuring β-galactosidase or or 4 days post infection by measuring luciferase activities. The left panels show the mean enzyme activities ± standard deviation derived from triplicate infections. RLU/s: relative light units per second. Middle panels show normalized infection rates in which reporter enzyme activities obtained from infected cells in the absence of inhibitor were set at 100%. The right panels depict the calculated IC50 values. The number above the bar represents the fold-change in the IC50 derived from 0.05 ng semen-exposed relative to 0.05 or 0.5 ng mock-exposed virus infection. Ns, no statistically significant difference; **** p<0.0001; *** p<0.001 (unpaired t-test).

Western blot analysis of XIAP and cIAP1 expression and PARP cleavage in H460, A549 and 34LU cells pre-treated with vorinostat or entinostat for 6h followed by the SMAC mimetic birinapant for 24h.

Human PBMCs containing indicated concentrations of Tenofovir disoproxil fumarate were inoculated with 5 ng mock-exposed or semen-exposed R5-HIV-luciferase, or 50 ng R5-HIV-luciferase as infectivity matched control. Infection rates were determined 3 days post inoculation.

Nystatin, filipin III, and oseltamivir inhibit the labeling of trophozoites by CTXB and GM1 antibodies. (A) Trophozoites were treated with nystatin (27 μM), filipin III (7.6 μM), and oseltamivir (20 μM) for 30 min before labeling with Alexa Fluor-conjugated CTXB antibody (images a to d) and GM1 antibody (images a′ to d′). Myriocin (27 μM), a sphingolipid inhibitor, was used as a negative control (images e and e′). N, nucleus; PM, plasma membrane; F, flagella; VD, ventral disc. Bars, 5 μm. (B and C) Changes in intensity of control and inhibitor-treated trophozoites. For intensity measurements, cells were randomly selected from 10 to 15 fields from 3 to 5 separate experiments. Approximately 50 cells were considered for each condition and were analyzed using Zeiss Zen 2009 confocal software. One-way ANOVAs were performed to evaluate differences (means ± SDs) between the treatment and control groups. Mean intensities from 3 to 5 separate experiments are shown in panels B and C. Statistical significance was calculated using a one-way ANOVA test, followed by the Tukey (B) and Holm-Šídák (C) methods. *, P < 0.05; **, P < 0.01; ***, P < 0.001; NS, not significant.

BLI of NSG mice engrafted with BV173, treated with no therapy (control), plerixafor: 1 mg/kg IP daily, ESKM 100 ug twice weekly, and a combination of ESKM and plerixafor. (A) Logarithmic plot of BLI of leukemia growth measured weekly. Error bars are 5-95% confidence intervals. There was a small but not significant difference between ESKM and combination treated group. (B) End of therapy (day 34) BLI.

Lethally irradiated C57BL/6 recipient mice were injected with a 1:1 mixture of GFP+JAK2(V671F) and wild-type bone marrow cells. Five weeks later, mice were treated with vehicle (C), hydroxyurea (H; 30 mg/kg twice daily IP), ruxolitinib (R; 30 mg/kg twice daily oral gavage), BMN673 (B; 0.33 mg/kg IV), H+R, H+B, R+B, and H+R+B for 3 weeks. Percentage of GFP+JAK2(V617F) was measured in (panel B) bone marrow cells, (panel C) splenocytes, and (panel D) peripheral blood leukocytes; (panel E) number of GFP+JAK2(V617F) Lin−Sca1+c-Kit+ (LSK) cells per 106 bone marrow cells was calculated, too. *P < .05, **P < .05, and ***P < .05 when compared with control, single treatment, and double treatment, respectively, from 6 to 7 mice using the Student t test.

Immunofluorescence of NF-κB/p65 in HCT 116 and SW 1116 cells exposed to TNFα (25 ng/ml) with or without nafamostat mesilate(FLU) (100 μM). Scale bars: 20 μm.

(C) Double immunolabeling showed CK-labeled cells (green) co-expressed with 40, 6-diamidino-2-phenylindole (DAPI) (blue), scale bars are 50 μm.

Immunocytochemistry demonstrates NR4A1 nuclear location at control (non-chymase treated) in HL-1 cells (a). Chymase (2.5 ug/ml) treatment for 2 h induces NR4A1 (red) cytoplasmic translocation in HL-1 cells as well as myosin (green) disruption (panel b). Active chymase enters HL-1 cells and is prevented by Dynasore (c and d). There marked entry into HL-1 cell nuclei and cytoplasm after treatment with chymase (5 ug/ml) for 2 h that is prevented by pre-treatment with Dynasore. Lack of co-staining with caveolin 3 (green) demonstrates that chymase is not transported via caveolae. Dynasore prevents transferrin uptake in HL-1 cells.

NCI-H929 EV and miR-137 OE cells were treated with ATM activator Chloroquine Phosphate (CQ), specific ATM inhibitor KU-55933, and ATR inhibitor AZ20 for 12 hr. Immunoblotting showed the expression of p-ATM, p-Chk2, p-BRCA1, p-ATR and p-Chk1.

(B) Cleaved PARP, Bax and Bcl2 protein expression was evaluated by immunoblotting of KRAS mutant cells lysates after 48 h of honokiol (10, 20, 40, and 60 μM) treatment. ∗∗P < 0.01 and ∗∗∗P < 0.001 for comparison between control group and honokiol-treated group.

Verification of Hdac6 deletion in knockout MEFs. Expression of HDAC6 and acetylation of tubulin were analyzed by immunoblotting.

Dose-response curve of the SH5190, SH5190 R292K and the A(H1N1)pdm09 (2167) viruses in the presence of peramivir (C) using a chemiluminescent (NA-star) assay. The NA activity of the H7N9 or H1N1 viruses under increasing concentrations (0.03-1000 nM) of the NAIs was determined. Each data point represents the normalized mean ± SEM neuraminidase activity from three replicated wells. The experiments were repeated twice.

Dose-response curve of the SH5190, SH5190 R292K and the A(H1N1)pdm09 (2167) viruses in the presence of oseltamivir zanamivir (B) using a chemiluminescent (NA-star) assay. The NA activity of the H7N9 or H1N1 viruses under increasing concentrations (0.03-1000 nM) of the NAIs was determined. Each data point represents the normalized mean ± SEM neuraminidase activity from three replicated wells. The experiments were repeated twice.

UA and Q-VD-Oph attenuated TNF-α-induced unclear translocation of NF-κB (C) and P-STAT3(S727) (D) in BEAS-2B cells.

(A) BoDV-1 infection was measured by IFA. BoDV-1 P40 was detected with a primary monoclonal antibody (red), nuclei were stained with DAPI (blue), merged image (scale bars: 50 μm). Favipiravir: T-705

Immunofluorescence of NF-κB (p65) (200×). SK-OV-3 and A2780S cells were treated with different compounds (vehicle, 5 μM JSH-23, 5 μM SC75741, 5 μM CAPE, 4 μM MIL, 4 μM MIL combined with pre-treated 5 μM JSH-23, 4 μM MIL combined with pre-treated 5 μM SC75741, 4 μM MIL combined with pre-treated 5 μM CAPE) for 48 h. NF-κB(p65) was detected by a fluorescence microscopy. The nuclei were stained with Hoechst 33258.

C, SA-beta gal staining results of A549-LKB1 cells treated by trametinib (30 nmol/L), radiotherapy (2 Gy), and HCQ (50 μmol/L). Cells were treated by HCQ and/or trametinib 4 hours prior to radiotherapy. Drugs were washed out 24 hours after radiotherapy. Cells were incubated for additional 48 hours before staining. D, Clonogenic survival assay of A549-LKB1 cells treated with trametinib (30 nmol/L) and HCQ (50 μmol/L).

(B) 2-DG activates AMPK and suppresses H3K27me3, PRC2 of ovarian cancer cells in normoglycemic condition.

(A) HCT 116 cells were treated with Tram (0, 30, or 60 nM), and then treated with 10 μM FTI-277/4 μM GGTI-298 or vehicle control for 48 hr. CCK8 assays were performed to assess cell proliferation and growth. The data from a representative example of three independent experiments are presented as the mean±SD. (B) SW480 and MDA-MB-231 cells were treated with Tram (0, 80, 160 nM), and then treated with 10 μM FTI-277/4 μM GGTI-298 or vehicle control for 48 hr. CCK8 assays were performed to assess cell proliferation and growth. The data from a representative example of three independent experiments are presented as the mean±SD.

Cell viabilities with increasing concentrations of cisplatin (CP) and doxorubicin (DOXO) under normoxic and hypoxic condition for 48 hours were determined by MTT assay. IC50 values are presented as the means ?SDs (n=4) and * denotes p<0.05.

BLI of NSG mice engrafted with BV173, treated with no therapy (control), plerixafor: 1 mg/kg IP daily, ESKM 100 ug twice weekly, and a combination of ESKM and plerixafor. (A) Logarithmic plot of BLI of leukemia growth measured weekly. Error bars are 5-95% confidence intervals. There was a small but not significant difference between ESKM and combination treated group. (B) End of therapy (day 34) BLI.

Same experiment as in Figure 2 with fluorescent daptomycin shown as confocal images. A 7/3 DOPC/DOPG GUV (including 1% Rh-PE) was introduced at time zero into a solution containing 1 uM daptomycin with 0.5 uM BODIPY-daptomycin and 1 mM Ca2+. To show colocalization of lipid and peptide in aggregates, confocal images were taken: red for RH-PE and green for BODIPY-daptomycin. As the protrusion length decreased, aggregates appeared on the GUV surface containing both lipid and peptide. The scale bar is 10 um.

CHX blocked Pim-1 AUG isoform translocation from the cytosol to mitochondria and led to reduced pBADS112. Cox-2 was examined to assess the effect of tigecycline on mitochondrial translation. Cy-Cox-2: cytoplasmic Cox-2; Mt-Cox-2: mitochondrial Cox-2.

(E) Left panel, Western blot showed that 20 μM Ribavirin effectively inhibits elevated Sox2 expression in irradiated SW1990 cells. Right panel, qPCR analysis showed that Sox2 mRNA level in irradiated SW1990 cells at 48 hours was lower than that in non-irradiated SW1990 cells, *p < 0.05. (F) Left panel, Western blot showed 20 μM Ribavirin inhibits elevated Sox2 expression in irradiated BxPc-3 cells. Right panel, qPCR analysis showed that there is no significant fluctuation of Sox2 mRNA in irradiated BxPc-3 cells and Ribavirin did not influence sox2 mRNA level in irradiated BxPc-3 cells.

(A) HONE1 and (B) HK1-EBV cells were treated with TPA (40 ng/ml) and SB (3 mM) for EBV induction. After 3 h, uninduced and induced HONE1 and HK1-EBV cells were treated with the vehicle control (0.006% DMSO) or indicated concentrations of berberine for 48 h. Subsequently, the culture medium was harvested for quantitative-PCR analysis with EBNA1 primers. Each sample was normalized to the amount of the GAPDH gene; *p<0.05, compared with the control group.

Schistosomicidal effects of PZQ, ART(Artemether) and OXA on 14-day schistosomula. Magnification of the in vitro-cultured schistosomula treated with PZQ, ART, and OXA at 10 μM. Drugs were added at day 14 and schistosomula were analyzed at day 21 (late treatment). 0.1% DMSO in medium (solvent of the drugs; used as negative control) had no effect on schistosomula development.

Effect of FDA approved drugs on EBOV transfected donor and recipient immune cells. (A) Varying concentrations of Cambinol and Esomepraole (1 nM, 1 μM, 10 μM), Oxytetracycline (1 nM, 10 nM, 10 μM), and DMSO (0.00001, 0.0001, 0.001%) treatments were administered to VP40-transfected 293T cells, followed by incubation for 3 days. Supernatant was centrifuged to remove cellular material and subsequently incubated for 72 h at 4°C with 20 μL of 30% slurry of NT80/82 beads. The NT pellet was washed twice with PBS and resuspended in SDS Laemmli buffer, followed by Western blot analysis for levels of exosomal markers CD63 and Alix. Actin levels were also analyzed. Mid-log phase EVTR2C cells (containing VP40) were treated with either low (0.1 μM), medium (1 μM), or high (10 μM) concentrations of Oxytetracycline and incubated for 5 days along with untreated 293T cells. Cell-free supernatants were harvested, filtered, and used to treat CEM (B), Jurkat (C), and U937 (D) cells. Cells were incubated for 5 days and assayed with CellTiter-Glo for viability. Statistical significance was determined using two-tailed student’s t-test with all groups compared to the untreated 293T cell control groups. RLU, Relative Luminescent Units. ∗p < 0.05, ∗∗∗p < 0.001.

Schistosomicidal effects of PZQ, ART and OXA on 14-day-schistosomula. Magnification of the in vitro-cultured schistosomula treated with PZQ, ART, and OXA at 10 μM. Drugs were added at day 14 and schistosomula were analyzed at day 21 (late treatment). 0.1% DMSO in medium (solvent of the drugs; used as negative control) had no effect on schistosomula development.

BALB/c mice, orally infected with E. multilocularis eggs, were treated by either mefloquine (100 mg/kg twice per week, n = 9), ABZ (200 mg/kg, 5 times per week, n = 8) or control-treated (placebo, n = 9). After 12 weeks of treatment, parasite lesion numbers in the liver were assessed microscopically (A).

Values represent the average scores for each group ± S.E. (N = 10 per group). The differences between individual experimental groups and 17 days DSS only group were statistically significant (p < 0.05) for all groups.

AmpC β-lactamase and MDH inhibition dose-response curves in the absence (black circles) and presence (red triangles) of DTT for (C and D) physcion.