ATPase

(F) Detection of expression of SDHA and TOMM20 by immunoblotting in wild-type and Atg7-/- K562 cells with indicated drugs. The expression levels were quantified with number indicated. (G) Quantification analysis of mitochondrial mass in wild-type and Atg7-/- K562 cells when cultured with oligomycin(100 ng/ml), antimycin A(50 μM) and Baf-A1(10 nM).

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.

(C) MyoIIA inhibition by blebbistatin treatment in vivo impairs leukemia cell extravasation into the brain. One hour before B-ALL cell transfer, C57BL/6 CD45.1+ recipient mice were treated with vehicle or blebbistatin (2.5 mg/kg). Twenty-four hours after B-ALL cell transfer, the recipient mice were intravenously injected with CD19-APC antibodies and then euthanized 4 min later.

Mitochondrial depolarization mediated relocation of mitochondrial RC-TPP into lysosomes and the ensuing lysosomal acidity triggered rhodamine fluorescence. RC-TPP-loaded Tom20-GFP+ HeLa cells were treated without or with CCCP (20 μM) in the presence or absence of BFA (200 nM). Colocalization of Tom20-GFP (in green) and mitochondrial RC-TPP (in blue) is shown in cyan. Scale bar: 10 μm.

CSFV multiplication was inhibited by GCA. (A) The safe concentration of GCA was tested using the MTT assay. The operations and calculations were the same as those used for BFA. (B) Cellular CSFV RNA with or without 400 nM GCA were examined using real-time PCR. (C) The titers of CSFV with or without GCA were tested by IFA. (D) CSFV proliferation in ST cells with or without GCA was observed by IFA at 24, 48 and 72 hpi. Results from the three independent experiments are shown as means ± SD. *P<0.05 and **P<0.01 compared with the control group.

Changes of mitochondrial membrane potential (ΔΨm). (A) Representative micrographs captured by confocal laser scanning microscope were evaluated for ΔΨm by staining with JC-1 at 24h post ionizing radiation. Red dimers indicated normal mitochondrial function and green monomers indicated collapse of ΔΨm with cytoplasmic fluorescence. Scale bar = 50μm. (B) Kinetics of changes of ΔΨm was showed as the ratio of red to green fluorescence in each treatment staining with JC-1 at 0.5, 6, 12, 24, 48, 72 and 96 h after X-ray radiation (IR) with or without 10μM BTB treatment. Data are expressed as means±SD of triplicate measurements. IR compared with IR+BTB, *p< 0.05, **p< 0.01, t-test. Control compared with IR+BTB, #p< 0.05, ##p< 0.01, t-test.

LN229 GBM cell lysates were immunoblotted with antibodies recognizing specific focal adhesion proteins. Note the decreased levels of Cas, paxillin, and FAK tyrosine phosphorylation in cells treated with CB-5083. In addition, CB-5083 treatment leads to decreased levels of total PTP-PEST expression, but Vcp protein expression levels are not impacted.

PQ increases glucose transport and the translocation of glucose transporters. Cells were treated with PQ for 48 h. The survival of cells treated with PQ in the presence or absence of phlorizin (e) was determined. Bar graph represents percents of viable cells (cell survival).

The changes in 10-N-nonylacridineorange (NAO) fluorescence indicated the cardiolipin (CL) oxidation. In normal cells, NAO could interact with nonoxidized cardiolipin and produces a characteristic green fluorescence. However, after cardiolipin is oxidized, NAO cannot bind to it. F-H, The changes in MDA, GSH, and SOD levels. I and J, The cytoplasmic [Ca2+] map via confocal microscopy by Fluo-2. Fluorescence intensity of Fluo-2 was measured by excitation wavelengths of 340 nm and emission wavelengths of 500 nm. Data (F/F0) were obtained by dividing fluorescence intensity (F) by (F0) at the resting level (t = 0), which was normalized to normal groups. Data are shown as means ± SEM. *P < .05 vs normal group; #P < .05 vs PA group, @P < .05 vs PA + siRNA-NR4A1 or PA + melatonin groups

Normal and Mutation human iPSC-CMs Structure when Treated with Omecamtiv Mecarbil. Human iPSC-CMs were plated on 100 kPa flat PDMS and grown for five days and left untreated, treated acutely on day 5, or treated continuously from the time of plating with the myosin activator Omecamtiv Mecarbil. iPSC-CMs were then fixed and stained for actin filaments (phalloidin, red) and nucleus (DAPI, blue). (a, c, e) Normal and (b, d, f) Mutation untreated, acutely treated, and continuously treated iPSC-CMs.

E, Analysis of BrCSC viability upon treatment with tamoxifen (tam) and fulvestrant (fulv) at the indicated doses. The experiments were performed in triplicates. ** P<0.01.

(F) Detection of expression of SDHA and TOMM20 by immunoblotting in wild-type and Atg7-/- K562 cells with indicated drugs. The expression levels were quantified with number indicated. (G) Quantification analysis of mitochondrial mass in wild-type and Atg7-/- K562 cells when cultured with oligomycin(100 ng/ml), antimycin A(50 μM) and Baf-A1(10 nM).

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.

(C) MyoIIA inhibition by blebbistatin treatment in vivo impairs leukemia cell extravasation into the brain. One hour before B-ALL cell transfer, C57BL/6 CD45.1+ recipient mice were treated with vehicle or blebbistatin (2.5 mg/kg). Twenty-four hours after B-ALL cell transfer, the recipient mice were intravenously injected with CD19-APC antibodies and then euthanized 4 min later.

Mitochondrial depolarization mediated relocation of mitochondrial RC-TPP into lysosomes and the ensuing lysosomal acidity triggered rhodamine fluorescence. RC-TPP-loaded Tom20-GFP+ HeLa cells were treated without or with CCCP (20 μM) in the presence or absence of BFA (200 nM). Colocalization of Tom20-GFP (in green) and mitochondrial RC-TPP (in blue) is shown in cyan. Scale bar: 10 μm.

CSFV multiplication was inhibited by GCA. (A) The safe concentration of GCA was tested using the MTT assay. The operations and calculations were the same as those used for BFA. (B) Cellular CSFV RNA with or without 400 nM GCA were examined using real-time PCR. (C) The titers of CSFV with or without GCA were tested by IFA. (D) CSFV proliferation in ST cells with or without GCA was observed by IFA at 24, 48 and 72 hpi. Results from the three independent experiments are shown as means ± SD. *P<0.05 and **P<0.01 compared with the control group.

Changes of mitochondrial membrane potential (ΔΨm). (A) Representative micrographs captured by confocal laser scanning microscope were evaluated for ΔΨm by staining with JC-1 at 24h post ionizing radiation. Red dimers indicated normal mitochondrial function and green monomers indicated collapse of ΔΨm with cytoplasmic fluorescence. Scale bar = 50μm. (B) Kinetics of changes of ΔΨm was showed as the ratio of red to green fluorescence in each treatment staining with JC-1 at 0.5, 6, 12, 24, 48, 72 and 96 h after X-ray radiation (IR) with or without 10μM BTB treatment. Data are expressed as means±SD of triplicate measurements. IR compared with IR+BTB, *p< 0.05, **p< 0.01, t-test. Control compared with IR+BTB, #p< 0.05, ##p< 0.01, t-test.

LN229 GBM cell lysates were immunoblotted with antibodies recognizing specific focal adhesion proteins. Note the decreased levels of Cas, paxillin, and FAK tyrosine phosphorylation in cells treated with CB-5083. In addition, CB-5083 treatment leads to decreased levels of total PTP-PEST expression, but Vcp protein expression levels are not impacted.

PQ increases glucose transport and the translocation of glucose transporters. Cells were treated with PQ for 48 h. The survival of cells treated with PQ in the presence or absence of phlorizin (e) was determined. Bar graph represents percents of viable cells (cell survival).

The changes in 10-N-nonylacridineorange (NAO) fluorescence indicated the cardiolipin (CL) oxidation. In normal cells, NAO could interact with nonoxidized cardiolipin and produces a characteristic green fluorescence. However, after cardiolipin is oxidized, NAO cannot bind to it. F-H, The changes in MDA, GSH, and SOD levels. I and J, The cytoplasmic [Ca2+] map via confocal microscopy by Fluo-2. Fluorescence intensity of Fluo-2 was measured by excitation wavelengths of 340 nm and emission wavelengths of 500 nm. Data (F/F0) were obtained by dividing fluorescence intensity (F) by (F0) at the resting level (t = 0), which was normalized to normal groups. Data are shown as means ± SEM. *P < .05 vs normal group; #P < .05 vs PA group, @P < .05 vs PA + siRNA-NR4A1 or PA + melatonin groups

Normal and Mutation human iPSC-CMs Structure when Treated with Omecamtiv Mecarbil. Human iPSC-CMs were plated on 100 kPa flat PDMS and grown for five days and left untreated, treated acutely on day 5, or treated continuously from the time of plating with the myosin activator Omecamtiv Mecarbil. iPSC-CMs were then fixed and stained for actin filaments (phalloidin, red) and nucleus (DAPI, blue). (a, c, e) Normal and (b, d, f) Mutation untreated, acutely treated, and continuously treated iPSC-CMs.

E, Analysis of BrCSC viability upon treatment with tamoxifen (tam) and fulvestrant (fulv) at the indicated doses. The experiments were performed in triplicates. ** P<0.01.