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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: PPP1R26 drives hepatocellular carcinoma progression by controlling glycolysis and epithelial-mesenchymal transition

Fig. 6

Overexpressed PPP1R26 induces accumulation of nuclear PKM2 and promotes EMT in HCC cells dependent on nuclear PKM2. A The expression of E-cadherin, Vimentin, Snail1 and Twist2 was determined by Western blot in the indicated cells (left). Relative mRNA levels of CDH1, VIM, SNAIL1 and TWIST2 were evaluated by RT-qPCR in the indicated cells (middle and right). Beta-actin was used as a loading control. B Western blot analysis of indicated protein levels when GFP-PPP1R26 was expressed in the PKM2-depleted Huh7 cells. C GFP-PPP1R26 (upper) or GFP (lower) was transfected into HepG2 and Huh7 cells. Cells were cultured under serum starvation and immunofluorescence staining was performed with anti-PKM2 antibody (red). DAPI was used to stain the nucleus. D HepG2 and Huh7 cells were transfected with GFP or GFP-PPP1R26 and cultured under serum starvation. Cytosolic and nuclear fractionations were prepared and the expression of PKM2 was determined by Western blot. TOP1 and Tubulin were used as the nuclear marker and cytoplasm marker, respectively. E Cells were treated with DMSO or TEPP-46 for 24 h after GFP-PPP1R26 (upper) or GFP (lower) was transfected. Immunofluorescence staining was done with anti-PKM2 (red). DAPI stained nuclei. Immunofluorescence images were taken using a confocal microscope. F Huh7 cells were treated with DMSO or TEPP-46 for 24 h after GFP-PPP1R26 was transfected. Whole cell extracts were used to evaluate the expression of GFP-PPP1R26 and E-cadherin by Western blot (upper). Relative mRNA levels of CDH1 after treatment with DMSO or TEPP-46 for 24 h in GFP-PPP1R26 or GFP expressing Huh7 cells (lower). Data information: In (A & F), data are presented as mean ± SD. Statistical significance was assessed using one-way ANOVA with post hoc analysis LSD test (A) or two-tailed t-tests (F). **P < 0.01 and ****P < 0.0001. ns denotes no significance

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