alda1  (MedChemExpress)

Journal: Journal of diabetes investigation

Article Title: Mitochondrial ALDH2 improves ß-cell survival and function against doxorubicin-induced apoptosis by targeting CK2 signaling.

doi: 10.1111/jdi.14230

Figure Lengend Snippet: Figure 2 | ALDH2 activation inhibits doxorubicin induced mitochondrial dysfunction. (a) The INS-1 cells were treated with doxorubicin (1 lM) with or without ALDA1 (25 lM) for 12 h (*P < 0.001 vs Control; **P < 0.005 vs DOXO). (b) AKT phosphorylation was quantified by immunoblotting. (c, d) The INS-1 cells were treated with doxorubicin (1 lM) with or without ALDA1 (25 lM) for 12 h (*P < 0.001 vs Control; **P < 0.005 vs DOXO). (e) cytochrome-c, cleaved caspase-3, and DNA damage marker cH2AX protein levels were quantified by immunoblotting. (f) Acid sphingomyelinase protein levels were quantified by immunoblotting. (g) The INS-1 cells were treated with C2-ceramide (50 lM) with or without ALDA1 (25 lM) for 8 h (*P < 0.005 vs Control; **P < 0.001 vs C2-CER). (h–j) Measurement of mitoROS and mitochondrial membrane potential in INS-1 cells after C2-ceramide (50 lM) treatment with or without ALDA1 (25 lM) for 8 h (*P < 0.005 vs Control; **P < 0.001 vs STZ; C2-CER). (j) Cytosolic cytochrome c and cleaved caspase-3 protein levels were quantified by immunoblotting. (k) GPX4 protein levels were measured through immunoblotting. (l) Measurement of lipid peroxidation in ALDA1 (25 lM) treated cells (*P < 0.005 vs Control; **P < 0.001 vs DOXO). (m, n) Cells were treated as mentioned above, and cell viability was analyzed by CCK assay (*P < 0.005 vs Control; **P < 0.001 vs C2-CER; DOXO).

Article Snippet: Reagents were sourced commercially as follows: Doxorubicin, ALDA1, C2-Ceramide (Sigma-Aldrich, MO, USA; K); and CX-4945 from SelleckChem.

Techniques: Activation Assay, Control, Phospho-proteomics, Western Blot, Marker, Membrane

Journal: Journal of diabetes investigation

Article Title: Mitochondrial ALDH2 improves ß-cell survival and function against doxorubicin-induced apoptosis by targeting CK2 signaling.

doi: 10.1111/jdi.14230

Figure Lengend Snippet: Figure 3 | CK2 inhibition reverses ALDH2 protective effect in b-cells. (a) INS-1 cells were treated with doxorubicin (1 lM) along with ALDA1 (25 lM) for 12 h with or without the CK2 inhibitor CX-4945 (5 lM). (b–e) INS-1 cells were treated with C2-ceramide (50 lM) along with ALDA1 (25 lM) for 8 h with or without the CK2 inhibitor CX-4945 (5 lM) (*P < 0.001 vs Control; **P < 0.005 vs C2-CER; ***P < 0.001 vs CX-4945). (f-h) INS-1 cells were treated with doxorubicin (1 lM) along with ALDA1 (25 lM) for 12 h with or without the CK2 inhibitor CX-4945 (5 lM). The cleaved caspase-3, cH2AX and GPX4 protein levels were quantified by immunoblotting. Measurement of lipid ROS levels by C11 BODIPY dye (*P < 0.001 vs Control; **P < 0.001 vs DOXO; ***P < 0.001 vs CX-4945). (i, j) Cells were treated as mentioned above, and cell viability was analyzed by CCK assay (*P < 0.001 vs Control; **P < 0.005 vs C2-CER; DOXO ***P < 0.001 vs CX-4945). Data are represented as mean – SD of three independent experiments.

Article Snippet: Reagents were sourced commercially as follows: Doxorubicin, ALDA1, C2-Ceramide (Sigma-Aldrich, MO, USA; K); and CX-4945 from SelleckChem.

Techniques: Inhibition, Control, Western Blot