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. Author manuscript; available in PMC: 2012 Jun 1.
Published in final edited form as: Cancer Discov. 2011 Oct 17;1(7):608–625. doi: 10.1158/2159-8290.CD-11-0046

Figure 2. Suppression of CRKL decreases the proliferation of NSCLC cells that harbor 22q11.21 amplifications.

Figure 2

(A) Effect of CRKL suppression on relative proliferation of NSCLC cell lines and AALE cells. Top, Immunoblot of CRKL proteins in each cell line expressing control shRNAs or two distinct CRKL-specific shRNAs. Bottom, Cell proliferation 6 d after infection with indicated shRNAs for each cell line. Data represent mean + s.d of six replicate measurements.

(B) Effect of expression of shRNA-resistant CRKL on the proliferation of H1755 cells expressing CRKL-specific shRNAs. Top, Generation of a shRNA-resistant CRKL cDNA (CRKLsilent mutant). The 21-mer sequence targeted by shCRKL#1 is marked in red color. Middle, Immunoblot of CRKL 4 d after infection with indicated shRNAs in H1755 cells expressing a control vector or V5-epitope tagged CRKLsilent mutant. Bottom, Effect of CRKL suppression by CRKL-targeting shRNAs on the proliferation of H1755 cells expressing a control vector or CRKLsilent mutant. The shCRKL#2 and shCRKL#3 target the 3′UTR of endogenous CRKL mRNA.

(C) Effect of CRKL suppression on cleavage of PARP and Caspase-3 proteins in NSCLC cells. Immunoblots of PARP and Caspase-3 proteins in H1755 and HCC1833 cells expressing the indicated shRNAs.

(D) Effects of mutations on CRKL function in H1755 cells. Left, Schematic of CRKL mutants. Right, Effect of expressing wild-type or mutant CRKL on the proliferation of the H1755 cells expressing a CRKL-specific shRNA (shCRKL#3). Data represent mean + s.d. of six replicate measurements. * indicates p<0.0001 as compared to cells expressing a control vector and shCRKL#3.