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. Author manuscript; available in PMC: 2013 May 17.
Published in final edited form as: Oncogene. 2011 Sep 26;31(20):2593–2600. doi: 10.1038/onc.2011.428

Fig. 3. Validation of representative PKCε target genes in NSCLC cells.

Fig. 3

H358 cells expressing shRNA control (CTRL) or shRNA for PKCε (ε#1 and ε#2) were assayed for expression of key apoptosis-related genes. A) For qPCR, RNA was isolated using the QIAGEN RNeasy kit and reverse transcribed to cDNA using random hexamers and the TaqMan Reverse Transcription kit (Applied Biosystems). Real-time PCR assays were performed in a 7300 ABI PCR System (Applied Biosystems), using TaqMan Gene expression assays and TaqMan Universal master mix. Human 18S rRNA was used as an endogenous control for normalization. The relative levels of mRNA compared to control were calculated according to the ΔCt method. B) Western blots and the corresponding densitometric analyses were carried out essentially as previously described (Oliva et al., 2008). The following primary antibodies were used: anti-Bik, anti-Bak, anti-caspase 3, anti-cIAP2, anti-Bcl2, anti-Bcl-XL (Cell Signaling, 1:1000 dilution), and anti-β-actin (Sigma, 1:50,000 dilution). Densitometric analysis of 3 independent experiments is shown. Data are expressed as the mean ± S.E.M. (n=3). *, p<0.05; **, p<0.01.