Fig. 4: FMRP modulates the WNT pathways regulating the WNT5B and CTNNB1 mRNA metabolism.
From: FMRP modulates the Wnt signalling pathway in glioblastoma
A Enrichment analysis of known FMRP target mRNAs and mutated in GBM according to The Cancer Genome Atlas. Shown are pathways that are significantly enriched in the PANTHER database (FDR < 0.05). B Cluster heat map represents the log2 fold change (FC) for the 156 genes differentially expressed in all the three shFMR1 GSCs. Values on a log2 scale are color-coded as indicated on the right. C Left panel, representative western blot of total extract (input), FMRP immunoprecipitation (IP-FMRP) and mock immunoprecipitation (IP-IgG). FMRP and its interactor FXR2P are detected. Right panel, HPRT1, CTNNB1, and WNT5B mRNAs were quantified by RT-qPCR. Shown is the enrichment immunoprecipitation/total, relative to H3 mRNA (mean ± SEM, n = 3, *P < 0.05, **P < 0.01, Student’s t-test). HPRT1 mRNA was used as a negative control; CTNNB1 mRNA is also a well-known FMRP target and was used as a positive control. D The histogram represents WNT5B and CTNNB1 mRNA levels quantified in shNTC and shFMR1 #148, #163, and #169 GSC cell lines. HPRT1 mRNA was used as normaliser (mean ± SEM, n = 3, **P < 0.01, Student’s t-test). E WNT5B and CTNNB1 mRNA levels in CTRL siRNA (siCTRL) and FMR1 siRNA (siFMR1) T89G cells detected by RT-qPCR (mean ± SEM, n = 3, **P < 0.01, ***P < 0.001, Student’s t-test). F mRNA stability assay in CTRL siRNA (siCTRL) and FMR1 siRNA (siFMR1) T89G cells. RNA was isolated at the indicated time points after Actinomycin D treatment and the stability of WNT5B and CTNNB1 mRNAs was analysed by RT-qPCR (mean ± SEM, n = 3, ***P < 0.001, ****P < 0.0001, two-way ANOVA).
