Figure 3

Autophagy inhibition aggravated Chang liver cell necrosis in response to nutrient deprivation. Chang liver cells were cultured in complete medium or EBSS for 24 h in the absence or presence of 10 μM CQ. (A) Cell viability was measured by CCK8. Data were shown as mean ± SEM (n = 6; *: p < 0.05; **: p < 0.01). (B) Whole cell lysates of the EBSS and EBSS + CQ groups were subjected to detect the indicated antibodies by immunoblot assay. (C) The morphologies of cells were captured by a light microscope. (D, E and F) The representative images of Hoechst 33342/PI staining were shown at magnification × 400 (D). PI positive/ Hoechst strong positive cells with condensed nuclei were considered as apoptotic ones (white arrows), and PI strong positive/ Hoechst weak positive cells were regarded as necrotic ones (red arrows). The percentage of apoptosis (E) or necrosis (F) was shown. Data were shown as mean ± SEM (n = 3; *: p < 0.05; **: p < 0.01; ns: no significance).