Fig. 6. Gαi3 is crucial for the activation of Akt-mTOR in pancreatic cancer cells.

RNA-sequencing (RNA-seq) was employed to analyze the gene expression profile of priPC-1 cells following the knockdown of Gαi3 using shRNA (“sh-Gαi3-s1”) in contrast to cells treated with a non-specific scramble shRNA (“shC”). This comparison elucidated a set of differentially expressed genes (DEGs), as depicted in volcano plots (A). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the DEGs in sh-Gαi3-s1-expressing priPC-1 cells is shown (B). Western blot analysis was performed to examine the activation states of AKT-mTORC1 signaling in priPC-1 cells bearing the described genetic modifications (C, D). priPC-1 cells with“sh-Gαi3-s1” were engineered to express either a constitutively active Akt1 mutant (caAkt1, S473D) or a control vector (“Vec”), expression of the described proteins was assessed in these cells (E); Cells were then cultured for 24-72 h, cell proliferation (F) and migration (G) were measured by the indicated experiments, with results quantified. *P < 0.05 versus “shC”/“Cas9-C”/“Vec” group. ^#P < 0.05. The experiments depicted in this figure were replicated five times (n = 5, biological repeats), consistently yielding similar results.