Fig. 3.

Inhibition of Pol I transcription initiation at rDNA operons impairs the EMT program. All cell culture experiments performed in NMuMG cells ± TGFβ, ± CX-5461 treatments labeled in each panel. a Quantification of FUrd ± TGFβ and ± CX-5461; P < 0.003. b qRT-PCR of 45S (pre)-rRNA transcript ± TGFβ ± CX-5461: Control vs. TGFβ, P < 0.05; TGFβ vs. TGFβ  + CX-5461, P < 0.02. c Quantification of EdU ± TGFβ ± CX-5461, P < 0.002. d ChIP analysis of UBF and Snail1 binding to the rDNA promoter ± TGFβ ± CX-5461: UBF, P < 0.027; Snail1, P < 0.018. e Immunostaining of Vimentin (green), Phalloidin (green) and Snail1 (green) ± TGFβ ± CX-5461. f Quantification of Vimentin (Vim) and Snail1 immunofluorescence from e, P < 0.02. g Relative percent invasion from Boyden chamber invasion assay ± TGFβ ± CX-5461, P < 0.002. h Immunostaining of Rictor (green), Calnexin (red), Rictor/Calnexin (white arrows, yellow) and DAPI (blue) ± TGFβ ± CX-5461. i Immunostaining of Rictor (green) ± RNase A treatment. j qRT-PCR of Vimentin (Vim), Snail1 and Rictor mRNA expression, P < 0.02. Asterisk denotes t-test significance. Error bars ± SE, n = 3. Scale bar for all images = 50 µm