Fig. 4

HucMDEs promote autophagic flux in L-O2 cells. a, b L-O2 cells were transfected with ATG5 or ATG7 siRNAs and were then treated with HucMDEs for 48 h. ATG5, ATG7, and MAP 1LC3B expression levels were examined by Western blotting. c L-O2 cells were treated with HucMDEs, 3-MA, or HucMDEs + 3-MA for 48 h. The level of MAP 1LC3B was monitored by Western blotting. d L-O2 cells were treated with HucMDEs, Baf, or HucMDEs + Baf for 48 h. The level of MAP 1LC3B was examined by Western blotting. e L-O2 cells were transfected with ATG5 or ATG7 siRNAs and were then treated with HucMDEs or DMSO for 48 h, after which they were observed via fluorescent microscopy to evaluate GFP-MAP 1LC3 expression patterns. f L-O2 cells were treated with HucMDEs, Baf, or HucMDEs + Baf for 48 h, after which they were observed by fluorescent microscopy to evaluate the GFP-MAP 1LC3 expression patterns. g, h Percentages of cells with more than four GFP-LC3B dots were quantified of the indicated groups. All results are expressed as the mean ± SD (*P < 0:05; **P < 0:01; ***P < 0.001)