IL-32 b

1. Supplementary Figure 2. Effect of IL-32 and siRNA of IL-32 on melanoma, colon and prostate cancer cell growth and apoptosis as well as IL-32 expression. (A), To determine effects of the IL-32 expression level on melanoma, colon and prostate cancer cell growth, the cells were inoculated into 24-well plates (5 × 104 cells/well), and transfected with the IL-32 plasmid (0.1-0.4 g/well) in the absence or presence of siRNA of IL-32b. Cell growth was measured by direct counting after trypan blue staining. Control, untransfected cells. *, p<0.05 compared with the colon and prostate cancer cells transfected with vector.(B), Effect of IL-32 on apoptotic cell death in colon and prostate cancer cell growth. Cells were transfected with the vector or the IL-32 plasmid for 24 h. The apoptotic index was determined as the number of DAPI-stained, TUNEL-positive cells counted. The results are expressed as mean ± SD of three experiments with each experiment performed in triplicate. *, p<0.05 compared with the colon and prostate cancer cells transfected with vector. (C), Expression ofIL-32 was determined with specific antibody of IL-32b as described in material and Methods. The expression band representative of three experiments with each experiment performed in triplicate.

2. Supp. Fig. 2 SW620 PC3 120 120 100 100 Cell viability (% of control) * * * 80 80 # 60 60 # 40 40 20 20 0 0 50 # 50 # # # 40 40 30 30 A Melanoma 20 * 20 * 10 120 10 0 100 0 * Cell viability (% of control) * 80 # 60 40 20 - - 50 100 - - siCON (nM) - - 50 100 - - 0 siCON (nM) - - 50 100 - - - - - - 50 100 siIL-32 (nM) - - - - 50 100 siIL-32 (nM) - - - - 50 100 B IL-32 (0.4 g/well) IL-32 (0.4 g/well) IL-32 (0.4 g/well) # # Apoptotic rate (%) Apoptotic rate (%) Apoptotic rate (%) * C IL-32b IL-32b IL-32b b-actin b-actin b-actin 5 # 8 # # 4 6 # 5 # # Foldincrease 3 Foldincrease 4 4 * 2 * 2 Foldincrease 3 1 1 * 2 0 0 1 0 siCON (nM) - - 100 - siCON (nM) - - 100 - siCON (nM) - - 100 - siCON (nM) - - 100 - siCON (nM) - - 100 - siCON (nM) - - 100 - siIL-32 (nM) - - - 100 siIL-32 (nM) - - - 100 siIL-32 (nM) - - - 100 siIL-32 (nM) - - - 100 siIL-32 (nM) - - - 100 siIL-32 (nM) - - - 100 IL-32 (0.4 g/well) IL-32 (0.4 g/well) IL-32 (0.4 g/well) IL-32 (0.4 g/well) IL-32 (0.4 g/well) IL-32 (0.4 g/well)