Fig. 4: Xist RNA turnover is independent of the m6A nuclear reader protein YTHDC1.
From: m6A and the NEXT complex direct Xist RNA turnover and X-inactivation dynamics
a, Strategy for in-frame insertion of FKBP12F36V into YTHDC1. b, Western blot showing the protein size and level of YTHDC1 upon 0 or 2 h of dTAG-13 treatment. The tagged protein is indicated by the red arrow. TBP was used as a loading control. The blot is representative of three biologically independent experiments. c, Western blot showing the protein level of YTHDC1–FKBP12F36V for ChrRNA-seq samples shown in d. TBP was used as a loading control. d, Box plot showing the allelic ratio of X-linked genes (n = 263) from ChrRNA-seq analysis for YTHDC1 dTAG degron (left 4 samples) and published data on PCGF3/5 degron (right 4 samples) mES cells. The experimental design is as described in Fig. 1c. The red dashed line indicates allelic ratio at 0.5. Samples and conditions are indicated above and below, respectively. P values, as indicated, were calculated using a two-sided paired t-test. Two replicates were averaged. Center lines indicate the median, box limits indicate the first and third quartiles and whiskers indicate 1.5× the IQR. e, Bar plot showing the expression level of Xist from ChrRNA-seq analysis for samples and conditions described in d, with individual replicates represented as dots.
