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. 2019 Nov 19;48(2):862–878. doi: 10.1093/nar/gkz1092

Figure 2.

Figure 2.

RGG box of FMRP attenuates AGO association and subsequent degradation of MOV10-FMRP co-bound mRNAs. (A) (Left) Immunoblot (ib) of endogenous proteins encoded by mRNAs co-bound by FMRP–MOV10 in HEK293 cells expressing FMRP or FMRP-ΔRGG. eIF5B is the loading control. Vertical line denotes the proteins that are ‘protected’. (Right) Quantification of immunoblots (n = 3), error bars represent SD, *P < 0.05, ** P < 0.001 Student t-test. (B) RT-qPCR of IP’ed AGO- associated mRNAs in cells expressing WT FMRP or FMRP-ΔRGG, (n = 3), error bars represent SD, ** P < 0.001 Student t-test. Statistical significance is for all four protected RNAs, MAZ through HN1L. (C) (Top) AGO was IP’ed and examined for GFP-tagged FMRP and FMRP-ΔRGG (GFP). (Bottom) GFP tagged FMRP and FMRP-ΔRGG were IP’ed and examined for AGO. (D) Luciferase activity of the MAZ 3′ UTR in cells expressing the WT FMRP or FMRP-ΔRGG constructs, (n = 3), error bars represent SD, ** P < 0.001 Student t-test. All experiments were performed in triplicate, representative immunoblots are shown.