Calcium Channel

The ROCK inhibitors fasudil and Y27632 prevented SCP2 cell bone metastasis in nude mice (n = 10 per group). Shown are BLI images of bone metastases, IHC analyses of SMAD3 C-tail phosphorylation and PTHLH, osteoclast TRAP staining, and BLI quantitation.

Levetiracetam reduces DNA, synaptic, and neurite damage caused by Dox. (A) An example of cortical cultures pre-treated with 5 μ M Lev and then Dox (0.01 μ M, overnight), fixed and stained with antibodies against MAP2c. Scale bar is 20 μ m. (B) Cortical neurons at 28–32 DIV were pre-treated with 5 μ M Lev, and then a vehicle or with Dox (0.01 μ M for 3 days, or 0.1 μ M, overnight) was added. Neurons were fixed, and stained for MAP2c, synapsin, and with the Hoechst dye, and imaged. Note that red MAP2c staining also includes Dox's red fluorescence. (C) Synaptically developed primary cortical cultures at 28−32 DIV were pretreated with 5 mM Lev, and then a vehicle or with Dox (0.01 μM for 3 days, or 0.1 μM, overnight) was added. Neurons were then fixed, and stained with antibodies against γH2A.X and MAP2c, and with the Hoechst dye. Blue staining was used by the algorithm to identify and analyze γH2A.X.

The proportion of SP cells in HepG2 and HCCLM3 cell lines were 0.16% and 32.2%, which reduced to 0% and 0.28% in the presence of verapamil.

Manidipine decreases the accumulation of HCMV Early and Late proteins. HFFs were infected with HCMV AD169 and then treated with test compounds or DMSO as a control. NTZ-treated samples were also included as a control for inhibition of E and L protein expression. At the indicated times p.i., total cell extracts were prepared and analyzed by WB with anti-IEA, anti-UL44, and anti-UL99 antibodies. Tubulin immunodetection served as a loading control. Images are representative of three independent experiments.

Assessment of the role of Ca2+ channels during RSV replication. HEp-2 cells were infected with RSV strain long at an MOI of 0.1, and treated with calcium channel blockers nifedipine, nimodipine, and tetrandrine (A), at indicated concentrations. Supernatants were collected at 48 h postinfection and viral titers were determined by immunoplaque assay. Grey bars represent cytotoxicity of the compounds. The data presented were obtained from two independent experiments. Error bars represent the standard deviations from two independent experiments. NS, no differences at a significance level of 0.05.

Results of hit validation experiments for these four compounds for animals treated at L1 stage at four different concentrations (0.5, 5, 25, and 50 μM). The blue bars represent the values obtained from the original screen as reference points. Error bars are standard error of the mean. Statistical significance for all different doses for individual compounds was calculated with respect to the lowest concentration using two-tailed, t-test (nZ70 animals except n = 37 for Geld 25 μM). The statistical significance values are represented as P<0.05 (*) and P<0.005 (**).

The fold change of C-peptide content after octreotide, nicorandil and nifedipine treatment. Octreotide, nicorandil and nifedipine decreased insulin secretion in wild-type, heterozygous mutated and homozygous mutated cells.

Gentisic acid reduces interstitial and perivascular fibrosis in mice with transverse aortic constriction (TAC). Representative images of Masson's trichrome staining of the hearts of mice in the sham and TAC + vehicle, gentisic acid, bisoprolol, or diltiazem groups. Scale bar = 50 μm. A, Interstitial fibrosis and B, perivascular fibrosis are shown. Fibrotic areas are stained blue.

Antidepressant effects caused by systemic ethosuximide injection (200 mg kg−1, i.p.; c, d) in the FST(forced swim test) and SPT (sucrose preference test) 1 h after treatment.

BMMs were seeded in a collage I-coated plate and incubated with RANKL (100 ng/ml) and M-CSF (50 ng/ml) for 5 d. When mature osteoclasts were observed, they were digested by collagenase and seeded on bovine bone slices. The bone slices were treated with or without tetrandrine for an additional 2 d. Then, the cells were stained with rhodamine–phalloidin to observe the F-actin rings, and the areas of F-actin rings were calculated by Image-Pro Plus 6.0 software.

Western blot analysis of myocardium lysate at 4 weeks. Hypertrophic markers were tested in mouse cardiomyocyte cell line HL-1 Bay-K-8644.

(B-E) The % of GFP-positive cells upon compound treatment were determined for the indicated HIV-1 latency cell lines (B. J-LAT 6.3; C. JLAT 10.6; D. CA5; E. EF7). Results were normalized to DMSO control. Values represent the mean ± s.d. (n = 4-6). * p < 0.05, one-way ANOVA followed by Tukey’s multiplecomparison test

The ROCK inhibitors fasudil and Y27632 prevented SCP2 cell bone metastasis in nude mice (n = 10 per group). Shown are BLI images of bone metastases, IHC analyses of SMAD3 C-tail phosphorylation and PTHLH, osteoclast TRAP staining, and BLI quantitation.

Levetiracetam reduces DNA, synaptic, and neurite damage caused by Dox. (A) An example of cortical cultures pre-treated with 5 μ M Lev and then Dox (0.01 μ M, overnight), fixed and stained with antibodies against MAP2c. Scale bar is 20 μ m. (B) Cortical neurons at 28–32 DIV were pre-treated with 5 μ M Lev, and then a vehicle or with Dox (0.01 μ M for 3 days, or 0.1 μ M, overnight) was added. Neurons were fixed, and stained for MAP2c, synapsin, and with the Hoechst dye, and imaged. Note that red MAP2c staining also includes Dox's red fluorescence. (C) Synaptically developed primary cortical cultures at 28−32 DIV were pretreated with 5 mM Lev, and then a vehicle or with Dox (0.01 μM for 3 days, or 0.1 μM, overnight) was added. Neurons were then fixed, and stained with antibodies against γH2A.X and MAP2c, and with the Hoechst dye. Blue staining was used by the algorithm to identify and analyze γH2A.X.

The proportion of SP cells in HepG2 and HCCLM3 cell lines were 0.16% and 32.2%, which reduced to 0% and 0.28% in the presence of verapamil.

Manidipine decreases the accumulation of HCMV Early and Late proteins. HFFs were infected with HCMV AD169 and then treated with test compounds or DMSO as a control. NTZ-treated samples were also included as a control for inhibition of E and L protein expression. At the indicated times p.i., total cell extracts were prepared and analyzed by WB with anti-IEA, anti-UL44, and anti-UL99 antibodies. Tubulin immunodetection served as a loading control. Images are representative of three independent experiments.

Assessment of the role of Ca2+ channels during RSV replication. HEp-2 cells were infected with RSV strain long at an MOI of 0.1, and treated with calcium channel blockers nifedipine, nimodipine, and tetrandrine (A), at indicated concentrations. Supernatants were collected at 48 h postinfection and viral titers were determined by immunoplaque assay. Grey bars represent cytotoxicity of the compounds. The data presented were obtained from two independent experiments. Error bars represent the standard deviations from two independent experiments. NS, no differences at a significance level of 0.05.

Results of hit validation experiments for these four compounds for animals treated at L1 stage at four different concentrations (0.5, 5, 25, and 50 μM). The blue bars represent the values obtained from the original screen as reference points. Error bars are standard error of the mean. Statistical significance for all different doses for individual compounds was calculated with respect to the lowest concentration using two-tailed, t-test (nZ70 animals except n = 37 for Geld 25 μM). The statistical significance values are represented as P<0.05 (*) and P<0.005 (**).

The fold change of C-peptide content after octreotide, nicorandil and nifedipine treatment. Octreotide, nicorandil and nifedipine decreased insulin secretion in wild-type, heterozygous mutated and homozygous mutated cells.

Gentisic acid reduces interstitial and perivascular fibrosis in mice with transverse aortic constriction (TAC). Representative images of Masson's trichrome staining of the hearts of mice in the sham and TAC + vehicle, gentisic acid, bisoprolol, or diltiazem groups. Scale bar = 50 μm. A, Interstitial fibrosis and B, perivascular fibrosis are shown. Fibrotic areas are stained blue.

Antidepressant effects caused by systemic ethosuximide injection (200 mg kg−1, i.p.; c, d) in the FST(forced swim test) and SPT (sucrose preference test) 1 h after treatment.

BMMs were seeded in a collage I-coated plate and incubated with RANKL (100 ng/ml) and M-CSF (50 ng/ml) for 5 d. When mature osteoclasts were observed, they were digested by collagenase and seeded on bovine bone slices. The bone slices were treated with or without tetrandrine for an additional 2 d. Then, the cells were stained with rhodamine–phalloidin to observe the F-actin rings, and the areas of F-actin rings were calculated by Image-Pro Plus 6.0 software.

Western blot analysis of myocardium lysate at 4 weeks. Hypertrophic markers were tested in mouse cardiomyocyte cell line HL-1 Bay-K-8644.

(B-E) The % of GFP-positive cells upon compound treatment were determined for the indicated HIV-1 latency cell lines (B. J-LAT 6.3; C. JLAT 10.6; D. CA5; E. EF7). Results were normalized to DMSO control. Values represent the mean ± s.d. (n = 4-6). * p < 0.05, one-way ANOVA followed by Tukey’s multiplecomparison test