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. 2006 Mar 27;172(7):983–990. doi: 10.1083/jcb.200508064

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Copyright © 2006, The Rockefeller University Press

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Figure 2. — Overexpression or down-regulation of SCCAs significantly affected UV-induced apoptosis. (A) FACS analyses of apoptotic cells using SCCA1- or SCCA2-transfected 3T3/J2 cells, 48 h after UV irradiation (30 mJ/cm²). Cells were stained with FITC-conjugated Annexin V and propidium iodide. (B) Analyses of five experiments are summarized. (C) 12 clones whose expression levels of SCCA1 mRNA distributed from 1 to 2,772-fold were established. Using these clones, the effects of UV irradiation were examined. Cells were harvested 48 h after UV irradiation (50 mJ/cm²) and FACS analyses were performed. The antiapoptotic activity correlated with SCCA1 expression. r = 0.734. (D) Using pSilencer vector, an siRNA construct targeted to a homologous sequence of SCCAs was stably transfected into HaCaT keratinocytes. Typical FACS analyses of nonirradiated (UV−) and UV-irradiated (UV+) siSCCA/HaCaT cells were shown. Apoptotic cells were analyzed 48 h after UV irradiation (75 mJ/cm²). (E) Statistical analyses of five experiments. Error bars represent the mean of five wells ± SD.