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Fig. 3 | Molecular Cancer

Fig. 3

From: Regulation of Sox2 and stemness by nicotine and electronic-cigarettes in non-small cell lung cancer

Fig. 3

a Depletion of E2F1, YAP1 or α7 nAChR by siRNA prevents the nicotine-mediated induction of Sox2. Depletion of Oct4 or TEAD2 did not have any impact on the induction. b A schematic showing the location of the E2F binding sites that were tested by ChIP assays; the location of forward and reverse primers for each site are indicated by arrows. c A ChIP assay shows the association of E2F1 with all the tested binding sites; E2Fs 2 and 3 were mainly associated with the binding site spanned by primers F2 and R2. There was no E2F associated with c-Fos promoter. d Nicotine stimulation induces the association of E2F1 with sites spanned by primers F1R1 and F2R2; surprisingly, Rb could also be detected on site F1R1. e A transient transfection experiment conducted on A549 cells showing the induction of Sox2-Luc reported by E2F family members. The data represented is with ± standard deviation (SD) values derived from three independent experiments. The statistical comparison was carried out by unpaired two tailed Student’s t-test. f YAP1 and E2F1 can induce the Sox2 promoter, and can show an additive effect in transient transfection experiments. The data represented is with ± standard deviation (SD) values derived from three independent experiments. The statistical comparison was carried out by one-way ANOVA. g Nicotine and E-cigarette extracts induce YAP1 levels in A549 cells and human mesenchymal stem cells, as seen by an immunofluorescence experiment

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