Figure 8. Mitochondrial targeting of... | Rockefeller University Press

Figure 8.

$Figure 8. Mitochondrial targeting of Bnip3FL isoform in Panc-1 is increased in Panc-1 cells deficient for Bnip3Δex3 variant. (A) Mitochondrial targeting of Bnip3FL is increased in Panc-1 cells during hypoxia after knockdown of Bnip3Δex3 isoform. Western blot analysis of mitochondrial and cytoplasmic fractions under normoxic and hypoxic conditions in the absence and presence of Bnip3Δex3 knockdown is shown; the filter was probed with a murine antibody directed against mitochondrial protein VDAC1, or cytosolic protein GAPDH was used to verify the integrity and purity of the cell fractionation. Bnip3FL was detected with a murine antibody directed against Bnip3. (B) ROS production in panc-1 cells after Bnip3Δex3 knockdown with siRNA-Bnip3Δex3 under hypoxia conditions. Epifluorescence microscopy of Panc-1 cells monitored for ROS production by DHE (red fluorescence). Bars, 40 µm. (C) Cell viability. Representative epifluorescence images of cells stained with vital dyes calcein-AM and ethidium homodimer-1 to visualize live (green) and dead (red) cells for conditions shown in B are shown. Bars, 40 µm. (D) Histogram for quantitative data shown in C. Data were obtained from at least n = 3–4 independent experiments counting ≥500 cells from n = 3 glass coverslips for each condition tested. Data are expressed as mean ± SD (error bars). *, statistically different from control (CTRL).$

Mitochondrial targeting of Bnip3FL isoform in Panc-1 is increased in Panc-1 cells deficient for Bnip3Δex3 variant. (A) Mitochondrial targeting of Bnip3FL is increased in Panc-1 cells during hypoxia after knockdown of Bnip3Δex3 isoform. Western blot analysis of mitochondrial and cytoplasmic fractions under normoxic and hypoxic conditions in the absence and presence of Bnip3Δex3 knockdown is shown; the filter was probed with a murine antibody directed against mitochondrial protein VDAC1, or cytosolic protein GAPDH was used to verify the integrity and purity of the cell fractionation. Bnip3FL was detected with a murine antibody directed against Bnip3. (B) ROS production in panc-1 cells after Bnip3Δex3 knockdown with siRNA-Bnip3Δex3 under hypoxia conditions. Epifluorescence microscopy of Panc-1 cells monitored for ROS production by DHE (red fluorescence). Bars, 40 µm. (C) Cell viability. Representative epifluorescence images of cells stained with vital dyes calcein-AM and ethidium homodimer-1 to visualize live (green) and dead (red) cells for conditions shown in B are shown. Bars, 40 µm. (D) Histogram for quantitative data shown in C. Data were obtained from at least n = 3–4 independent experiments counting ≥500 cells from n = 3 glass coverslips for each condition tested. Data are expressed as mean ± SD (error bars). *, statistically different from control (CTRL).