Figure 6. siRNA mediated knockdown of CHRNB4 in lung cancer cell lines.

H1299 and A549 cells were treated with 20 nM of a siRNA pool against the CHRNB4 transcript for 96h. Cells were seeded in quadruplicates. Cell proliferation was measured employing A) HOECHST staining which represents the amount of DNA and B) with Calcein staining which measures the metabolic activity in a cell population. C) Expression of CHRNB4 in knockdown cells was detected by qPCR and normalized to expression in siCON treated cells. D) A time course experiment with Hoechst measurement every 24h revealed diverging growth curves for cells treated with siRNAs against CHRNB4 compared to those ones that were treated with siCON siRNAs E) and F) Deconvolution experiments with four single siRNAs (20nM) in H1299 and A549 G) Knockdown for both cell lines in the deconvolution experiment H) CHRNB4 knockdown cells show a reduced propensity to form colonies. Results are displayed from a 6-well plate and also in a bar chart.

Signal intensity is given as raw values (relative units)

*: p<0.05

**: p<0.01

***: p<0.001

Ns: not significant