Figure 8.
SNX10 is important for mitochondrial bioenergetics. (A) Mitochondrial oxygen consumption rate (OCR) was assessed in control and SNX10 knocked down cells using the Seahorse XFe24 Analyzer. OCR was measured following sequential addition of oligomycin, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), and rotenone/antimycin A (Rot/AntiA). (B) The four basal OCR measurements per well were averaged to determine the basal OCR value, and non-mitochondrial respiration was subtracted to ascertain the basal respiration associated with each condition. (C) ATP production was calculated by subtracting the proton leak from the maximal respiratory capacity. Error bars represent the mean ± SEM from n = 5. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple comparison test. Data distribution was assumed to be normal but was not formally tested. (D) CS activity was determined by spectrophotometry from lysates of U2OS cells transfected with siRNA for 72 h, in the presence or absence of DFP for the last 24 h. The graph displays mean values normalized to siCtrl. Significance was determined from n = 3 independent experiments by two-way ANOVA followed by Tukey’s multiple comparison test. Data distribution was assumed to be normal but was not formally tested. (E and F) Expression levels of CS were measured in control (siCtrl) and SNX10 knockdowns (siSXN10#1 and siSXN10#2) across three independent experiments. Band densities of CS were normalized to the housekeeping gene GAPDH. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test to compare each knockdown group to the control group. Data distribution was assumed to be normal but was not formally tested. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, and **** = P < 0.0001; nonsignificant differences are not depicted. Source data are available for this figure: SourceData F8.

SNX10 is important for mitochondrial bioenergetics. (A) Mitochondrial oxygen consumption rate (OCR) was assessed in control and SNX10 knocked down cells using the Seahorse XFe24 Analyzer. OCR was measured following sequential addition of oligomycin, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), and rotenone/antimycin A (Rot/AntiA). (B) The four basal OCR measurements per well were averaged to determine the basal OCR value, and non-mitochondrial respiration was subtracted to ascertain the basal respiration associated with each condition. (C) ATP production was calculated by subtracting the proton leak from the maximal respiratory capacity. Error bars represent the mean ± SEM from n = 5. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple comparison test. Data distribution was assumed to be normal but was not formally tested. (D) CS activity was determined by spectrophotometry from lysates of U2OS cells transfected with siRNA for 72 h, in the presence or absence of DFP for the last 24 h. The graph displays mean values normalized to siCtrl. Significance was determined from n = 3 independent experiments by two-way ANOVA followed by Tukey’s multiple comparison test. Data distribution was assumed to be normal but was not formally tested. (E and F) Expression levels of CS were measured in control (siCtrl) and SNX10 knockdowns (siSXN10#1 and siSXN10#2) across three independent experiments. Band densities of CS were normalized to the housekeeping gene GAPDH. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test to compare each knockdown group to the control group. Data distribution was assumed to be normal but was not formally tested. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, and **** = P < 0.0001; nonsignificant differences are not depicted. Source data are available for this figure: SourceData F8.

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