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. 2012 Mar 12;109(13):4980–4985. doi: 10.1073/pnas.1111596109

Fig. 4.

Fig. 4.

RNAi knockdown of SLC7A2-IT1A/B transcripts. Human neuroblastoma Kelly cells were transiently transfected with 10 nM of a nontargeting siRNA pool (control siRNA) or a 10 nM pool of four SLC7A2-IT1-targeting siRNAs (Pool). The targeting siRNAs were further tested individually at 10 nM each. In the figure “30” is A-specific, “589” is specific for A and B transcripts, “1326” is preAluSz-specific, and “1497” is L1PA8 specific. (A) Real-time PCR analysis to estimate SLC7A2-IT1A/B transcript levels. Values were normalized to relative β-actin levels. (B) Detection of active Caspase-3 protein levels by Western blotting. A representative original blot is shown. β-Actin served as loading control. (C) Statistical analysis of Western blotting results. Knockdown of SLC7A2-IT1A/B by ∼40% leads to a more than 2.5-fold elevated active Caspase-3 level. n = 3. (D) Correlation of SLC7A2-IT1A/B transcript levels and active Caspase-3 protein levels. Using different transfection conditions (10 nM up to 100 nM siRNA concentrations for 24 h up to 48 h) SLC7A2-IT1A/B transcript levels were assessed by real-time PCR and correlated to corresponding active Caspase-3 protein levels. PCC, Pearson correlation coefficient; n = 32. *P < 0.05, **P < 0.01, ***P < 0.001.