Figure 3

Suppression of NFAT2 expression by siRNA attenuates CORT-induced expression of FasL. A. After cells were transfected with the siRNA, NFAT2 siRNA394 or NFAT2 siRNA1232, targeting NFAT2 gene for 48 h, the expression of NFAT2 mRNA was analyzed by RT-PCR with primers specific to NFAT2. The cells transfected with scrambled siRNA served as negative control, and beta-actin mRNA served as internal controls. Lane1. 100-bp marker; Lane 2. Scrambled siRNA; Lane3. NFAT2 siRNA394; Lane 4. NFAT2 siRNA1232. B. Cells were transfected with the scrambled siRNA or siRNA targeting NFAT2 gene for 72 h, and expression of NFAT2 protein was analyzed by Western blotting with specific antibody. Lane1. Scrambled siRNA; Lane 2. NFAT2 siRNA394; Lane 3. NFAT2 siRNA1232. C. Expression of FasL mRNA was determined by RT-PCR in wild mLTC-1 (a) and mLTC-1 transfected with NFAT2 siRNA394 (b) after cells were treated with CORT or CORT plus CsA. Lane 1. Marker; Lane 2, 3 and 4 is without CORT treatment, with CORT treatment and CORT plus CsA treatment, respectively. D. Expression of FasL protein was determined by Western blotting in wild mLTC-1 (a) and mLTC-1 transfected with NFAT2 siRNA394 (b) after cells were treated with CORT or CORT plus CsA. Lane 1, 2 and 3 is without CORT treatment, with CORT treatment and CORT plus CsA treatment, respectively.