Journal:

Article Title: Protein Kinase C- α and ERK1/2 Mediate Mitochondrial Dysfunction, Decreases in Active Na + Transport, and Cisplatin-induced Apoptosis in Renal Cells

doi: 10.1074/jbc.M206373200

Figure Lengend Snippet: The monolayers were treated and RPTC functions analyzed as described under “Experimental Procedures.” White columns, controls; black columns, 50 μm cisplatin; light gray columns, 50 μm PD98059 + 50 μm cisplatin; dark gray columns, 10 nm Go6976 + 50 μm cisplatin; hatched columns, 3 μm UO126 + 50 μm cisplatin; striped columns, 50 μm zVAD-fmk + 50 μm cisplatin. Results are the average ± S.E. of three to five independent experiments (RPTC isolations).

Article Snippet: PKC- α inhibitor (Go6976) and MEK inhibitors (PD98059 and UO126) were supplied by Calbiochem (La Jolla, CA). zVAD-fmk was purchased from Biomol (Plymouth Meeting, PA).

Techniques:

Journal: Redox Biology

Article Title: Superoxide induced inhibition of death receptor signaling is mediated via induced expression of apoptosis inhibitory protein cFLIP

doi: 10.1016/j.redox.2019.101403

Figure Lengend Snippet: Death receptor sensitization upon GW is associated with decrease in intracellular O 2 •- . (A) CEM/Neo and CEM/Bcl-2 cells were pretreated for 1hr with 50 μM ZVAD-fmk before triggering apoptosis with anti-Fas (0.25 μg/ml for 18 hrs) in culture medium or in GW conditions and PI staining was used to assess DNA fragmentation (sub-G1 fraction) as described in Materials and Methods. (B) Cells were incubated for 4 hrs with 0.25 μg/ml of anti-Fas in normal medium or GW conditions and caspase 8 and (C) caspase 9 and 3 activities were determined in whole cell lysates using fluorometric assays that detect cleavage of specific substrates, as described in Materials and Methods. Enzyme activity are shown as fold increase over the untreated cells. (D) Cells were pretreated with PMA (62.5 ng/ml) for 1hr before treatment for 4 hrs with anti-Fas (0.25 μg/ml) in normal cell culture medium or GW medium and apoptosis was assessed by PI staining (sub-G1 fraction). (E) Caspase 8 activity was determined in the lysates from cells treated with anti-Fas in the presence and absence of PMA using a fluorometric assay and presented as fold increase over untreated cells. (F) Processing of capase-3 and 8 was determined by Western blot analysis using specific antibodies as described in Materials and Methods. Where applicable, data are Mean ± S.D. of three independent experiments and P values (* <0.05; ** <0.01; *** <0.005) were calculated by Ordinary one-way ANOVA using GraphPad Prism.

Article Snippet: Death receptor sensitization upon GW is associated with decrease in intracellular O 2 •- . (A) CEM/Neo and CEM/Bcl-2 cells were pretreated for 1hr with 50 μM ZVAD-fmk before triggering apoptosis with anti-Fas (0.25 μg/ml for 18 hrs) in culture medium or in GW conditions and PI staining was used to assess DNA fragmentation (sub-G1 fraction) as described in Materials and Methods. (B) Cells were incubated for 4 hrs with 0.25 μg/ml of anti-Fas in normal medium or GW conditions and caspase 8 and (C) caspase 9 and 3 activities were determined in whole cell lysates using fluorometric assays that detect cleavage of specific substrates, as described in Materials and Methods.

Techniques: Staining, Incubation, Activity Assay, Cell Culture, Western Blot