Skip to main content
Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: UBL4A inhibits autophagy-mediated proliferation and metastasis of pancreatic ductal adenocarcinoma via targeting LAMP1

Fig. 3

UBL4A-induced inhibition of tumor depends on autophagy. a Western blotting analyses of LC3B and p62 in four PDAC cell lines of different groups (Vector, LV-UBL4A-Flag, shCtrl, LV-shUBL4A). b Electron microscopy showed the different account of autophagosomes in SW1990 and CFPAC-1 transfected with lentiviral vector encoding UBL4A (LV-UBL4A-Flag) (bar, 100 nm). c Whole SW1990 and CFPAC-1 lysates in empty vector, LV-UBL4A-Flag, Chloroquine (CQ), LV-UBL4A-Flag plus CQ were subjected to western blotting to test E-cadherin, N-cadherin, p62, LC3B and UBL4A protein expression. d Whole PANC-1 and BxPC-3 lysates in shCtrl, shUBL4A, CQ, shUBL4A plus CQ were subjected to western blotting to test E-cadherin, N-cadherin, p62, LC3B and UBL4A protein expression. e-g The role of CQ in UBL4A-induced migration and invasion was demonstrated by transwell assay in SW1990 and PANC-1 (original magnification, 10×) (bars, 25 μm). h-j Wound healing assay was performed to detected the role of CQ in UBL4A-mediated metastasis in SW1990 and PANC-1 (original magnification, 10×) (bars, 25 μm). k-m The immunofluorescence assays were performed in pancreatic cancer cells that were transfected with flag-tagged mRFP-GFP-LC3 lentiviral vector in four different groups (original magnification, 20×) (bars, CFPAC-1: 25 μm, PANC-1: 10 μm). 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

Back to article page