Fig. 4

RP11-296E3.2 regulates the transcription and translation of STAT3 but not JAK2. A KEGG enrichment analysis of the SDEGs (99 overlapping) between sh-Con- and sh-RP11-296E3.2-transduced RKO cells. P. adjust, hypergeometric test with the Benjamini–Hochberg correction. B GSEA of the whole 45,009 genes between in sh-Con- and sh-RP11-296E3.2-transduced RKO cells. The P value was determined by a hypergeometric test. This analysis was performed by GSEA4.3.2 software. C mRNA levels of 5 genes in the JAK/STAT3 pathway from the RNA-seq results. D, F GEPIA2 analysis of the correlations between RP11-296E3.2 and JAK2 or STAT3 expression. E, G The altered mRNA levels of JAK2 and STAT3 were confirmed by qRT-PCR in HCT116, RKO, and SW48 cells transduced with the indicated shRNAs (n = 3 per group). H WB analysis of JAK2, p-JAK2, STAT3 and p-STAT3 in HCT116, RKO and SW48 cells transduced with the indicated shRNAs (n = 3 per group). I The STAT3 promoter was evaluated by a luciferase reporter assay in RP11-296E3.2-overexpressing and RP11-296E3.2-silenced cells. The data are presented as the means ± SDs. Statistical significance was assessed using one-way ANOVA followed by Tukey’s test for multiple comparisons. All experiments were performed with at least three biological duplicates (n = 3) for each group. *P < 0.05, **P < 0.01, ***P < 0.001