caspase 12 inhibition  (R&D Systems)

Journal: Journal of Biomedical Science

Article Title: A neuronal death model: overexpression of neuronal intermediate filament protein peripherin in PC12 cells

doi: 10.1186/1423-0127-19-8

Figure Lengend Snippet: The activity of calpain, caspase-12, caspase-9, and caspase-3 is increased on 8 day of NGF induction in pEGFP-Peripherin cells compared to PC cells . (A) Western blot analysis showing increased activation of caspase-12 (42 kD), caspase-9 (40 kD and 38 kD), and caspase-3 (19 kD and 17 kD) on later stage of NGF induction in pEGFP-Peripherin cells compared to PC cells. (B-E) Density of the bands for the calpain BDP (B), active caspase-12 (C), active caspase-9 (D), or active caspase-3 (E) relative to the β-actin band. The values are presented as the mean ± SEM for three experiments in each group. *p < 0.05, **p < 0.01 vs. control PC12 cells.

Article Snippet: When used, the selective calpain inhibitor calpeptin (20 μM) (Calbiochem, La Jolla, CA), the caspase-9 inhibitor Ac-LEHD-CMK (20 μM) (Calbiochem), the caspase-12 inhibitor Z-ATAD-FMK (2 μM) (MBL, Nagoya, Japan) was added to the medium on day 6, with 1% DMSO being used as the vehicle control.

Techniques: Activity Assay, Western Blot, Activation Assay

Journal: Journal of Biomedical Science

Article Title: A neuronal death model: overexpression of neuronal intermediate filament protein peripherin in PC12 cells

doi: 10.1186/1423-0127-19-8

Figure Lengend Snippet: Effects of calpain, caspase-9, and caspase-12 inhibitors on the survival of pEGFP-Peripherin cells . pEGFP-Peripherin cells were treated for 48 h with 20 μM calpeptin (calpain inhibitor), 20 μM Ac-LEHD-CMK (caspase-9 inhibitor), or 2 μM Z-ATAD-FMK (caspase-12 inhibitor) starting on day 6 of NGF induction. (A) Proteins were analyzed by Western blotting. Caspase-3 activity was detected by levels of the 120 kD caspase-3 breakdown product of α-fodrin (caspase-3 BDP). UT = untreated group. Equal loading of protein was confirmed using β-actin. (B) Cell viability was evaluated using the XTT assay. The data are presented as the mean ± SEM for eight experiments in each group. ** p < 0.01 versus untreated group. (C) The mitochondria membrane potential was evaluated by the ability of the cells to take up the fluorescent dye TMRE. Data are presented as the mean ± SEM for twelve experiments in each group. * p < 0.05 versus the untreated group.

Article Snippet: When used, the selective calpain inhibitor calpeptin (20 μM) (Calbiochem, La Jolla, CA), the caspase-9 inhibitor Ac-LEHD-CMK (20 μM) (Calbiochem), the caspase-12 inhibitor Z-ATAD-FMK (2 μM) (MBL, Nagoya, Japan) was added to the medium on day 6, with 1% DMSO being used as the vehicle control.

Techniques: Western Blot, Activity Assay, XTT Assay