Mitochondrial Dysfunction and Apoptosis in Retina

The retinas were collected at 1, 3, 5, and 7 days after the intravitreal injection of mtDNA or control solution. The retinas processed with the Mitochondria Isolation Kit for Tissue as previously described [4 (link)] and the proteins collected from the supernatant and sediment were used for the detection of cytochrome c. Western blotting was performed according to methods described in previous study [4 (link)]. The following primary antibodies were used: rabbit anti-BAX (#2772, Cell Signaling Technology), rabbit anti-BAK (#12105, Cell Signaling Technology), rabbit anti-caspase 9 (AF6348, Affinity Biosciences Ltd), rabbit anti-cleaved caspase 9 (AF5240, affbiotech), rabbit anti-caspase 3 (ab44976, Abcam), rabbit anti-cleaved caspase 3 (ab49822, Abcam), anti-β-actin (ab8227, Abcam), anti-cGAS (ab179785, Abcam), rabbit anti-STING (D1V5L) (#50,494, Cell Signaling Technology), rabbit anti-TBK1/NAK (D1B4) (#3504, Cell Signaling Technology), rabbit anti-phospho-TBK1/NAK (Ser172) (D52C2) XP^® (#5483, Cell Signaling Technology), rabbit anti-IRF3 (D83B9) (#4302, Cell Signaling Technology), rabbit anti-phospho-IRF3 (Ser396) (D6O1M) (#29047, Cell Signaling Technology), anti-interferon β (ab140211, Abcam), rabbit anti-cytochrome c (10993-1-AP, Proteintech), and rabbit anti-VDAC1 (ab154856; Abcam). Three eyes from each group were tested.

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Guo Y., Gan D., Hu F., Cheng Y., Yu J., Lei B., Shu Q., Gu R, & Xu G. (2022). Intravitreal injection of mitochondrial DNA induces cell damage and retinal dysfunction in rats. Biological Research, 55, 22.

Publication 2022

rabbit anti-BAK AF6348 ab44976 ab49822 ab8227 ab179785 ab140211 ab154856

Actin Antibodies Caspase 3 Caspase 9 Cgas Cytochrome c Eyes Interferon β Irf3 Isolation Mitochondria Mtdna Proteins Rabbit Retinas Tissue Vdac1

Corresponding Organization :

Other organizations : Eye & ENT Hospital of Fudan University

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Variable analysis

independent variables

Intravitreal injection of mtDNA or control solution

dependent variables

Cytochrome c detection
Levels of BAX, BAK, caspase 9, cleaved caspase 9, caspase 3, cleaved caspase 3, cGAS, STING, TBK1/NAK, phospho-TBK1/NAK, IRF3, phospho-IRF3, interferon β

control variables

Retinas collected at 1, 3, 5, and 7 days after injection
Retinas processed using Mitochondria Isolation Kit for Tissue

controls

Positive control: Retinas collected after intravitreal injection of mtDNA
Negative control: Retinas collected after intravitreal injection of control solution

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