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. Author manuscript; available in PMC: 2019 Jul 12.
Published in final edited form as: Cell. 2018 Jun 21;174(2):391–405.e19. doi: 10.1016/j.cell.2018.05.043

Figure 4. LINE1 promotes translation and ES self-renewal independently of Dux.

Figure 4

(A) Colony-formation assay showing that Dux knockdown does not rescue self-renewal upon LINE1 KD. Data are mean +/− s.e.m., n=3 biological replicates.

(B–C) GSEA plot showing preferential downregulation of the KEGG ribosome pathway following LINE1 KD, with either co-transfection of (B) Control or (C) Dux siRNAs.

(D) RNA per cell in ES cells 48h after nucleofection with RC or LINE1 ASOs. Data are mean +/− s.e.m., n=6 independent batches of equal numbers of cells.

(E) Diagram of experiments labelling nascent RNA/proteins with EU/HPG following ASO nucleofection.

(F) Representative histogram (left) of nascent transcription in RC or LINE1 ASO-treated samples 24–48h after nucleofection, with cells incubated without EU shown as control, and quantification (right) showing the relative decrease in translation upon LINE1 KD. Data are = mean +/− s.e.m., n=2 independent experiments.

(G) qRT-PCR showing decrease in rRNA and ribosomal protein gene expression 48h after LINE1 KD, shown as mean +/− s.e.m, n=3 biological replicates.

(H) Representative histogram (left) and quantification (right) as in (G), but performed for HPG incubations 48h after LINE1 KD. Data are = mean +/− s.e.m., n=4 independent experiments. See also Figure S4.