Validation of Change in rRNA 2'O-Methylation Pattern by FMRP
(A) RTL-P for selected sites on 18S and 28S rRNA to show differences in methylation in Shef4 WT and Shef4 FMR1 KO hESC lysate (n = 3, unpaired Student's t test, mean ± SEM).
(B) Schematic for reverse transcription followed by PCR (RTL-P) on FMR1 knockdown or FMRP overexpression in HeLa cells.
(C) Representative immunoblots for FMRP showing its knockdown in HeLa cells with Tubulin as a control.
(D–F) RTL-P for selected sites showing the change in 2’O-methylation on FMR1 knockdown in HeLa cells (n = 3, unpaired Student's t test, mean ± SEM).
(G) Representative immunoblots for FMRP showing overexpression of FLAG-FMRP in HeLa cells with Tubulin as a control.
(H–J) RTL-P for selected sites showing the change in 2’O-methylation on overexpression of FLAG-FMRP in HeLa cells (n = 3, unpaired Student's t test, mean ± SEM).