Fig. 1: PA caused apoptotic death of astrocytes by autophagic induction. | Cell Death & Disease

Fig. 1: PA caused apoptotic death of astrocytes by autophagic induction.

From: The autophagic degradation of Cav-1 contributes to PA-induced apoptosis and inflammation of astrocytes

Fig. 1

a Western blots showing the protein levels of LC3B-II in the astrocytes incubated with PA (0.25 mM) for the indicated times in the absence and presence of CQ (50 μM) (mean ± S.E.M. n = 3, *P < 0.05 vs. control; #P < 0.05 vs. corresponding time without CQ). b Representative fluorescent images showing the LC3B puncta formation in astrocytes incubated with PA (0.25 mM, 12 h) in the absence and presence of CQ (50 μM). Scale bar, 25 μm. The LC3B puncta area per cell was analyzed by Image J software and the fold change in LC3B puncta area relative to the control was plotted (mean ± S.E.M. n = 3, *P < 0.05 vs. BSA; #P < 0.05 vs. PA). c Representative transmission electron microscopy images of the astrocytes incubated with PA for 12 h. Yellow arrows denote autophagosomes. Scale bar, 5 μm. The number of autophagosomes per cross-sectioned cell were calculated (mean ± S.E.M. n = 10/group, *P < 0.05 vs. BSA). d Western blots showing the expression of LC3B-II in the astrocytes exposed to PA (0.25 mM, 12 h) in the absence and presence of 3-MA or CQ (mean ± S.E.M. n = 3, *P < 0.05 vs. BSA; #P < 0.05 vs. PA). e The Annexin-V/PI labeling flow cytometry showing the apoptotic rate (Annexin V+/PI− and Annexin V+/PI+) and necrotic rate (Annexin V−/PI+) of astrocytes exposed to 0.25 mM PA in the absence and presence of 3-MA (10 mM) (mean ± S.E.M. n = 3/group, *P < 0.05 vs. BSA; #P < 0.05 vs. PA). f Western blots showing the expression of Fas, CASP3, cleaved CASP3, and Bax/Bcl-2 ratio in the astrocytes exposed to PA (0.25 mM, 12 h) in the absence and presence of 3-MA (10 mM) (mean ± S.E.M. n = 3, *P < 0.05, **P < 0.01 vs. BSA; #P < 0.05 vs. PA). g The Annexin-V/PI assay showing the apoptotic rate (Annexin V+/PI− and Annexin V+/PI+) and necrotic rate (Annexin V−/PI+) of ATG5 or ATG7-deficient cells incubated with PA (0.25 mM, 12 h) (mean ± S.E.M. n = 3/group, *P < 0.05, **P < 0.01 vs. BSA + SC-siRNA; #P < 0.05 vs. PA + SC-siRNA). h Western blots showing the expression of Fas, CASP3, cleaved CASP3, and Bax/Bcl-2 ratio in ATG5 or ATG7-deficient cells incubated with PA (0.25 mM, 12 h) (mean ± S.E.M. n = 3, *P < 0.05, **P < 0.01 vs. BSA + SC-siRNA; #P < 0.05 vs. PA + SC-siRNA)

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