Fig. 3.

Integrative neurobiology study reveals a role of OCIAD1 in mitochondria-associated cell death. (a) Workflow for predicting OD1 (OCIAD1) function. (b) Cell death pathway and mitochondria are predicted as functional targets for OD1. Predicted functional partners of OD1 are indicated as nodes relevant to cell death pathways (blue), to mitochondria (orange), or both (green). (c-d) OD1 in the mitochondria was visualized via dual immunofluorescence staining against mitochondrial protein marks (SOD2) or Ds-Red Mito (c), and compared between the cerebral cortex of transgenic Tg AD mice (APP/PS1-ΔE9) and wild type mice (WT, d) at 4 months. Anti-OD1 (green), anti-SOD2 or DsRed (red). (e) Workflow for in vitro validation of OD1 function in mitochondria-associated cell death. (f-h) OD1 modulation affects cell injury in primary neurons induced by Aβ₁-₄₂ (f), in HT22 cells by MG132 (g), staurosporine (STR), H₂O₂ or glutamate (h). Cells infected with control vector encoding GFP only (CON, Mock, pCDH, pSIH), or plasmid encoding OD1 and GFP (OD1 or pCDH-OD1), shRNAi against OD1 (pSIH-OD1). (See also Fig. S3a-3d). (i) Up-regulation or down-regulation (j) of OD1 modulates the intrinsic apoptosis induced by MG-132 in HT22 cells. (k-l) Elevating OD1 in HT22 cells facilitates Str-induced apoptosis and release of cytochrome C (Cytc, k), SMAC (cyto-SMAC), EndoG (cyto-EndoG), but does not affect Caspase-8 activation (CASP8, l). Control vector (Mock), plasmid encoding OD1 (OD1). MG-12 h, MG-132 treatment for 12 hrs; STR treatment for 0 hr (0H), 4 hrs (Str4H), 8 hrs (Str8H), or 12 hrs (STR-12H). Data are presented as mean±SEM in (d-g) and (i- l)(student t-test, n = 3). * P < 0.05 vs. WT in (d) and vs.Veh in (f) #P < 0.05, ##P < 0.01 vs. Con in (g-h); *P < 0.05, **P < 0.01 vs. Con in (i-j); *P < 0.05, **P < 0.01 vs. Mock (k) or 0H (l), #P < 0.05 vs. Mock (l). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)