Fig. 6

LAMP1 involves in UBL4A-mediated inhibition of autophagy. a, b SiLAMP1 was transiently transfected in UBL4A overexpressed cell lines (SW1990, CFPAC-1) and forty-eight hours after transfection, the cell lysates were subjected to western blotting. c, d LAMP1 plasmid was transiently transfected in UBL4A downregulated cell lines (PANC-1, BxPC-3) and the cell lysates were subjected to western blotting. e Representative fluorescent photographs of autophagy flux assay in mRFP-GFP-LC3 tagged CFPAC-1 were subjected to Vector, LV-UBL4A-Flag, siLAMP1, LV-UBL4A-Flag +siLAMP1 groups (original magnification, 20×) (bar, 200 μm). f Representative fluorescent photographs of autophagy flux assay in mRFP-GFP-LC3 tagged PANC-1 were subjected to control, lv-shUBL4A, LAMP1 plasmid and lv-shUBL4A + LAMP1 plasmid groups (original magnification, 20×) (bar, 20 μm). g The numbers of GFP and mRFP dots were determined by fluorescent puncta in 5 high-power fields The ratio of red dots (autolysosomes) to yellow dots (autophagosomes) per cell was calculated. The statistical significance between different groups was calculated with Student t-test. Data are shown as the mean ± SD of three replicates; *P < 0.05, **P < 0.01; ***P < 0.001; ns: not significant