Extended Data Fig. 10: NOL9, LAS1L, and XRN2 catalytic point mutations.
From: Rixosomal RNA degradation contributes to silencing of Polycomb target genes
a, Immunoblot showing protein levels in control siControl (siCtrl), siNOL9, siNOL9+NOL9 wild type expressing plasmid, and siNOL9+ NOL9-K312A expressing plasmids. Actin served as a loading control. b, Immunoblot showing protein levels in control siControl (siCtrl), siLAS1L, siLAS1L+LAS1L wild type expressing plasmid, and siLAS1L+ LAS1L-R155A/H160A (LAS1l-2A) expressing plasmids. GAPDH served as a loading control. c, Immunoblot showing protein levels in control siControl (siCtrl), siXRN2-1 (siRNA 1), siXRN2-2 (siRNA 2), siXRN2-1+XRN2 wild type expressing plasmid, and siXRN2-1+ XRN2 E203G expressing plasmids. GAPDH served as a loading control. d, RT-qPCR analysis of RNA levels of H2B-CTRN in the indicated siRNA-treated and NOL9-rescued HEK293FT cells after 21 days of Doxcyline treatment. RNA expression levels were normalized to ACTB, and every knockdown was normalized to siCtrl. Error bars represent standard deviations for three biological replicates. Data are presented as mean values +/− SEM.
