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Fig. 3 | BMC Cancer

Fig. 3

From: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

Fig. 3

RCC2-YFP interrupts apoptosis via blocking Rac1 activation. a Protein lysate from HeLa cells expressing YFP or RCC2-YFP were immunoprecipitated with anti-GFP antibody and Western blotted with anti-GFP, anti-Rac1, anti-cdc42, or anti-RhoA antibody. Rac1 was co-precipitated with RCC2-YFP. b HeLa cells expressing YFP or RCC2-YFP were serum-starve1d overnight followed by serum-stimulation for 5 min. GTP-bound activated Rac1 was pulled down by PAK-PBD beads and Western blotted with anti-Rac1 antibody. RCC2-YFP expression blocked both endogenous and serum-induced Rac1 activation. c HeLa, MDA-MA-231 and CRL5800 cells expressing YFP or RCC2-YFP were treated with STS (0.5 μM for HeLa; 10 μM for MDA and CRL5800) for 30 min and Western blotted with antibodies to phospho-JNK and total JNK. RCC2-YFP expression blocked the STS-induced JNK phosphorylation in all three cell lines. d Tumor cells transfected with Rac1-Q61L, RCC2-YFP or both for 48 h were treated with STS (0.5 μM for HeLa; 10 μM for MDA and CRL5800) and apoptotic cells were scored by DAPI stain after 24 h treatment. Rac1-Q61L expression in tumor cells revoked the protection of RCC2-YFP towards STS-induced apoptosis (mean ± S.D. of four replicates, *P < 0.05). e HeLa cells transfected with Rac1-Q61L, RCC2-YFP or both for 48 h in the presence of serum. Spontaneous apoptotic cells were scored by DAPI stain. Rac1-Q61L expression in HeLa cells revoked the protection of RCC2-YFP toward spontaneous apoptosis (mean ± S.D. of four replicates, *P < 0.01). f HeLa cells were transfected with Rac1-Q61L, RCC2-YFP or both in the presence of serum, and Caspase-Glo® 3/7 assay was performed 48 h after transfection. Rac1-Q61L and RCC2-YFP expression in HeLa cells affected Caspase 3/7 activity (mean ± S.D. of six wells, *P < 0.05)

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