Fig. 4: Screening and validation of the expression and interaction of the target gene SPTBN1. | Communications Biology

Fig. 4: Screening and validation of the expression and interaction of the target gene SPTBN1.

From: IGF2BP3 recruits NUDT21 to regulate SPTBN1 alternative polyadenylation and drive ovarian cancer progression

Fig. 4

A–C RT‒qPCR was used to detect the relative changes in the lengths of the target genes ICMT, ZEB1, RNF213 and SPTBN1 in A2780, OVCAR3, and SKOV3 cells following NUDT21 (A), IGF2BP3 (B) and METTL3 (C) knockdown (n = 4). D MeRIP‒qPCR of SPTBN1 m6A modification; RIP‒qPCR of SPTBN1 interaction with IGF2BP3 and NUDT21 (n = 4). E and F RIP-qPCR was used to validate the relative fold change in SPTBN1 long and short isoform binding with NUDT21 in OVCAR3 cells (F) following IGF2BP3 knockdown (E) (n = 4). G and H RIP-qPCR was used to validate the relative fold change in SPTBN1 long and short isoform binding with NUDT21 in OVCAR3 cells (H) following METTL3 knockdown (G) (n = 3). I A dual-luciferase assay was performed on the 32nd intron of SPTBN1 with WT and m6A site mutations, which were both cotransfected with IGF2BP3 (n = 4). J–L SPTBN1 long and short isoform protein levels were detected by Western blot in A2780, OVCAR3, and SKOV3 cells after NUDT21 (J), IGF2BP3 (K), and METTL3 (L) were knocked down. *P < 0.05, **P < 0.01, ***P < 0.001.

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