Oxidative Medicine and Cellular Longevity / 2019 / Article / Fig 5

Research Article

Icariin Inhibits AGE-Induced Injury in PC12 Cells by Directly Targeting Apoptosis Regulator Bax

Figure 5

Bax is a direct target protein for ICA. (a) The PC12 lysates were incubated with ICA beads or control beads, and then the potential proteins bound to the beads were resolved by SDS-PAGE, followed by silver staining. The asterisk-marked bands indicated nonspecific bead-bound vimentin and actin. (b) SPR analysis of ICA binding to Bax. The kinetic parameters of , , and were derived by fitting to a 1 : 1 Langmuir binding model. (c) ICA beads were incubated with PC12 lysates or recombinant Bax protein in the absence or presence of ICA for competitive binding, and the proteins bound to the beads were analyzed by western blot or silver staining. (d) ICA promoted Bax resistant to pronase (DARTS). PC12 lysates were incubated with pronase in the absence or presence of ICA, and Bax was detected by western blot. (e) PC12 lysates were incubated with ICA (20 μM) or vehicle followed by the cellular thermal shift assay (CETSA). Data were presented as from independent experiments performed in triplicate.

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