Fig. 5: TRIB3 is required for PIERCE1-mediated activation of the AKT pathway. | Oncogene

Fig. 5: TRIB3 is required for PIERCE1-mediated activation of the AKT pathway.

From: Impaired AKT signaling and lung tumorigenesis by PIERCE1 ablation in KRAS-mutant non-small cell lung cancer

Fig. 5

Relative mRNA (a) and protein (b) expression levels of TRIB3, pATK (S473), AKT, ACTIN, and FLAG 24 hours after transfection of empty vectors (EV) or PIERCE1-FLAG (P1-FLAG) in A549 cells. ACTIN was used as a loading control. c–e Relative mRNA levels of TRIB3 48 hours after siRNA-mediated transient KD (siP#1 and siP#3, c) or stable KD (shP#1 and shP#3, d) of PIERCE1, and in one-month old WT, PIERCE1 TG, and PIERCE1 KO mouse lungs (n = 3 per genotype. e ACTIN was used as a loading control (triplicated). Western blot analyses of TRIB3, actin, and FLAG in 1-month-old WT and PIERCE1 TG (f), and PIERCE1 KO (g) mouse lungs. h Western blot analyses for pAKT (S473), AKT, TRIB3, and ACTIN 48 hours after transient KD of indicated genes in A549 cells. i Monitoring of cell growth by crystal violet staining 72 hours after siRNA transfection against control (siCTL), PIERCE1 (siP1), and TRIB3 (siTRIB3) in A549 cells. j, k The ratios of PIERCE1 and TRIB3 expressions in lung cancer patients. Wachi Lung from Oncomine database (j) and The Cancer Genome Atlas (TCGA, k) database were used for the analyses. The r value indicates Pearson’s correlation coefficient. Error bars are ±SD *P < 0.05; **P < 0.01.

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