FIGURE 3.

bGP is inhibited in cells exposed to H₂O₂. a, HEK293T cells were transfected or not transfected with pCMV6-PYGB plasmid, and cells were exposed to 500 μm H₂O₂ for 20 min before being harvested. Cell lysis was then performed in the presence or absence of a reducing agent (10 mm DTT), and whole-cell extracts were assayed for endogenous glycogen phosphorylase activity. Data are expressed as the percentage of the control and represent mean values of three independent experiments ± S.D. ***, p < 0.001 when compared with positive control (upper panel); ###, p < 0.001 when two non-control groups are compared. Non-reduced and reduced whole-cell extracts were Western blotted and revealed for brain glycogen phosphorylase using an anti-bGP antibody. Ponceau red stains of the membranes are shown (lower panel). b, U87MG cells were exposed to 500 μm H₂O₂ for 20 min before being harvested. Cell lysis was then performed in the presence or absence of a reducing agent (10 mm DTT), and whole-cell extracts were assayed for endogenous glycogen phosphorylase activity. Data are expressed as the percentage of the control and represent mean values of three independent experiments ± S.D. ***, p < 0.001 when compared with positive control (upper panel); ###, p < 0.001 when two non-control groups are compared. Western blotting analysis of bGP from cells was revealed for brain glycogen phosphorylase using anti-bGP antibodies. Ponceau red stains of the membranes are shown (lower panel).