Journal: Communications Biology

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

doi: 10.1038/s42003-025-08097-6

Figure Lengend Snippet: 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.

Article Snippet: To produce shRNAs for lentiviruses, complementary sense and antisense oligonucleotides targeting METTL3 , IGF2BP3 , NUDT21 , and SPTBN1 were synthesized (Tsing Ke Biotechnology, China), annealed, cloned, and inserted into the pLKO.1 vector (Addgene).

Techniques: Knockdown, Modification, Binding Assay, Luciferase, Western Blot