Fig. 6
ZnONP-induced autophagy dysfunction in HaCaT cells. A Acridine orange staining was employed to examine the number of acidic vesicular organelles (AVOs), such as autophagolysosomes, in cells. Flow cytometry analysis demonstrated the effect of PT against ZnONP- and UVB-induced keratinocyte autophagolysosomes. The FL3 positive region (Q1 + Q2 regions) indicated the AO positive staining. B Histograms represent the percentage of acridine orange positive cells. C, D Western blot analysis of the effect of ZnONPs and UVB on the autophagy proteins LC3B and p62 in HaCaT cells. E Immunofluorescence staining with anti-LC3B/anti-LAMP1 antibodies in HaCaT cells treated with ZnONPs (10 μg/mL), UVB (68 mJ/cm2) + ZnONPs (10 μg/mL) or UVB (68 mJ/cm2) + ZnONPs (10 μg/mL) + PT (2 μM) for 24 h. Arrows indicate LC3B/LAMP1 co-localized puncta. Values are presented as the mean ± SD (n = 3). *p < 0.05, control group versus treatment groups. #p < 0.05, the UVB + ZnONPs groups versus the UVB + ZnONPs + PT groups