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. 2016 Jul 6;7(33):52832–52848. doi: 10.18632/oncotarget.10451

Figure 2. Gelsolin modulates SOD activity.

Figure 2

(A & B) Total intracellular SOD activity was measured from total cell lysates. Increased gelsolin expression lowers the SOD activity in both C1 and C8 cells as compared to wild type HCT116 and empty vector control cells. (B) Knockdown of endogenous gelsolin in HCT116 cells shows increased SOD activity when compared to the control siRNA treated cells. (C & D) Cytosolic and mitochondrial fractions were isolated in HCT116 cells following either gelsolin overexpression or gelsolin knockdown. Fractionation was confirmed by the presence of MnSOD and VDAC in the mitochondria and Cu/ZnSOD and GAPDH in the cytosol. β-actin was used as the internal loading control. (E & F) Cu/ZnSOD activity was measured from the cytosolic extract. Gelsolin overexpression results in suppression of Cu/ZnSOD activity when compared to the empty vector control cells whereas knockdown of gelsolin in HCT116 and C1 cells results in elevated Cu/ZnSOD activity. (G & H ) MnSOD activity was determined from the mitochondrial fraction. No significant change in MnSOD activity was observed when gelsolin was overexpressed or silenced in HCT116 and C1 cells. *p-value <0.05 versus controls using a two tailed Student's t-test. Values (mean ± SD ) are expressed as fold over the empty vector control or the control siRNA, which is arbitrarily set as one. The western blot gel pictures are representative images from three independent experiments.