Figure 3: Stard7 deficiency leads to excessive generation of ROS and mitochondrial DNA damage. | Scientific Reports

Figure 3: Stard7 deficiency leads to excessive generation of ROS and mitochondrial DNA damage.

From: The Phosphatidylcholine Transfer Protein Stard7 is Required for Mitochondrial and Epithelial Cell Homeostasis

Figure 3

(a) BEAS-2B and Stard7KDBEAS-2B cells were incubated with or without DCFDA (20 μM) and/or TBHP (25 μM) to induce oxidant stress and cultured for 3 hours. ROS were analyzed by flow cytometry. Data are expressed as MFI (mean ± SEM, n = 3); *p = 0.032, BEAS-2B cells vs. Stard7KDBEAS-2B cells; **p = 0.0186, TBHP-treated BEAS-2B cells vs. TBHP-treated Stard7KDBEAS-2B cells. (b) Knockdown cells were transiently transfected with Stard7-1HA or Stard7-2HA and ROS levels assessed as described in panel a. *p = 0.0111, Stard7KDBEAS-2B cells vs. BEAS-2B cells; **p = 0.0072, Stard7-1HA transfected Stard7KDBEAS-2B cells vs. Stard7KDBEAS-2B cells. (c) Relative mitochondrial DNA copy numbers were determined by qRT-PCR using primers for mitochondrial genes (16 sRNA, tRNA and Cox4) or a nuclear gene (actin). (d) Expression of the oxidant–sensitive gene interferon alpha inducible protein 27 (IFI27) was assessed in BEAS-2B and Stard7KDBEAS-2B cells by qRT-PCR before and after treatment with MitoTEMPO. MFI: mean fluorescent intensity. DCFDA: dichlorofluorescin diacetate; TBHP: tert-butyl Hydroperoxide.

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