Fig. 5

PLACT1 directly interacts with hnRNPA1. a PLACT1 sense and antisense RNAs were used in pull-down assays in PANC-1 cells, followed by electrophoresis and silver staining. HnRNPA1 is shown by a red arrow. b Mass spectrometry assays identified the PLACT1-interacting protein as hnRNPA1. c Western blotting analysis of proteins captured by PLACT1 sense and antisense fragments, indicating that PLACT1 associates with hnRNPA1. d RIP assays revealed that PLACT1 bound to hnRNPA1. e The colocalization of PLACT1 and hnRNPA1 was assessed by FISH and immunofluorescence. Scale bar: 5 μm. f qRT-PCR analysis indicated efficiency of hnRNPA1 knockdown and PLACT1 expressions in the hnRNPA1 knockdown cells. g-h Western blotting analysis showed the hnRNPA1 expression after PLACT1 overexpression (g) or knockdown (h) in PDAC. i-j EdU (i) and Transwell (j) assays revealed that depletion of hnRNPA1 partly reversed the effects of PLACT1-overexpressing PANC-1 cells. Representative images (left panel) and histogram analysis (right panel) are shown. Scale bars: 100 μm. p-values were calculated by using two-tailed t-tests and ANOVA followed by Dunnett’s tests for multiple comparison. The error bars represent standard deviations of three independent experiments. *p < 0.05 and **p < 0.01