Fig. 4

Regulation of NKILA expression in NSCLC cells. (a) qRT-PCR was performed to detect the relative NKILA levels in A549 and H226 cell cytoplasm and nucleus. GAPDH and NEAT1 RNA were used as fractionation indicators. (b) NKILA expression levels of A549 and H226 treated with TNFα, IL-1β or PBS were detected by qRT-PCR. (c) The schematic diagram of PMEPA1 and NKILA genome location. Exons are indicated by boxes, arrows indicate the direction of transcription. (d) The NKILA expression levels of A549 and H226 treated with TGF-β1 or PBS were detected by qRT-PCR. (e) NKILA expression kinetics in A549 and H226 cells following TGF-β1 stimulation. (f) Expression of NKILA assayed by qRT-PCR in A549 and H226 cells induced by TGF-β1 with or without TGF-β or NF-κB inhibition by SB505124 (SB) and JSH-23 (JSH) respectively. (g) Localization of Smad2/3 complex to NKILA promoter in A549 cells treated with TGF-β1 or PBS for 30 mins analyzed by ChIP. Data are expressed as means ± SEM, n = 3. Two-tailed Student’s t-test was used. ***p < 0.001