Fig. 2. Generation of inducible NAA40-KD colon cancer cell lines.

a Quantitative real-time PCR (qRT-PCR) analysis of NAA40 mRNA levels normalized to β-actin performed in HCT116, HT-29, SW480, and SW620 cell lines expressing doxycycline-inducible scramble shRNA (SCR), or two different shRNA sequences targeting NAA40 mRNA (NAA40-KD1 or NAA40-KD2). The relative values of NAA40 transcripts represent the mean ± s.d. of three independent experiments. Unpaired two-tailed Student’s t-test was used (**p < 0.01, ***p < 0.001). b Western blot analysis of cell extracts derived from dox-treated SCR or NAA40-KD HCT116, HT-29, SW480, and SW620 cells using antibodies against NAA40 and β-actin as loading control. The numbers below each blot indicate densitometry analysis of NAA40 protein levels in NAA40-KD relative to SCR after normalization to the β-actin bands