Figure 1.

Meteor transcriptional elongation promotes Eomes expression in naive mESCs

(A) Perturbations used to analyze the Meteor locus in this study. Shown are the locations of gRNAs used for deleting or inverting the Meteor promoter (pKO) or gene body (fKO) or for inhibiting transcription (dCas9-KD); the locations of insertion of ribozyme (Rz) or polyadenylation (pAS) sequences; and the scheme for knocking in the PGK promoter downstream of the Meteor promoter (pEX). The fKO and Rz-KD mESC lines are the same as in Tuck et al.³⁴

(B) RNA-seq tracks showing Meteor expression in the various pKO and pInv lines (grown in serum-free 2i/LIF conditions) and dCas9-KD lines (serum/LIF conditions). All tracks are normalized to the same scale. Orange denotes transcription on the plus strand, and blue denotes transcription on the minus strand.

(C) RNA-seq quantifications of Meteor and Eomes in Meteor pKO and pInv cell lines grown in serum/LIF (left) or serum-free 2i/LIF (right) conditions. Amounts normalized to WT1. Bars represent standard errors; n = 3. ^∗p < 0.05, ^∗∗p < 0.005.

(D) qRT-PCR quantifications of Meteor and Eomes levels in Meteor fKO and Rz-KD mESCs grown in serum-free 2i/LIF conditions. Levels were normalized to WT4 and Ppib for internal control. Bars represent standard errors; n = 8. ^∗p < 0.05, ^∗∗p < 0.005.

(E) Same as (D), for Meteor pKO and pInv mESCs grown in primed conditions, normalized to WT1. n = 4.

(F) Same as (D), for pAS clones grown in serum-free 2i/LIF conditions. Levels were normalized to WT7. n = 3.

(G) Same as (C), for Meteor dCas9-KD lines grown in serum/LIF conditions. Amounts normalized to Ctrl. n = 3. The dCas9-KD efficiencies of Meteor were 85% and 94% for KD1 and KD2, respectively.

(H) Same as (D), for pEX clones grown in serum-free 2i/LIF conditions. Levels were normalized to WT9. n = 3.

See also Figure S1.