Fig. 4

SOGA1 is specially recognized by IGF2BP1. A Silver staining revealed SOGA1-bound proteins from SW620 cells. B Immunoblotting of IGF2BP1/2/3, YTHDC1, YTHDF1/2/3 after RNA pull down assay with cell lysate (input), biotinylated-SOGA1 (case probe), and beads only (ctrl probe) in SW620 cells. C SOGA1 protein expression was detected in SW480 and SW620 cells with or without IGF2BP1 knockdown by western blotting. D SOGA1 mRNA expression was detected in SW480 and SW620 cells with or without IGF2BP1 knockdown by qRT-PCR. E, G SOGA1 protein expression in SW620 cells transfected with siRNAs of IGF2BP2, IGF2BP3 or YTHDF1 was examined by western blotting respectively. F SOGA1 mRNA expression was detected in SW620 cells with or without IGF2BP2 or YTHDF1 knockdown by qRT-PCR. (H, I) IGF2BP1 expression in the TCGA CRC cohort. J, K Association of IGF2BP1 mRNA expression with lymph node metastasis (J), and distant metastasis (K) in CRC patients in TCGA database. L Kaplan-Meier survival curves of OS based on IGF2BP1 expression in CRC patients in TCGA database. M RIP-qPCR displayed the relative enrichment of SOGA1 mRNA in each group precipitated with lgG or IGF2BP1 antibody with the normalization to input. IP efficiency of IGF2BP1 was validated using western blotting. GAPDH was used as protein control. N The capacity of IGF2BP1 binding to SOGA1 in SW480 cells was detected by RNA pulldown assay. O Stability of SOGA1 mRNA was detected in IGF2BP1-konckdown and control cells via qRT-PCR at the indicated time after actinomycin D (5 µg/ml) treatment. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001