Fig. 4

Huwe1 interacted with Gadd45b at 24 h after reperfusion. a, b Protein was extracted from normal neuron cells or neuron cells exposed to OGD/R 24 h, and prepared from CO-IP assay. Protein was incubated with IgG antibody, as a negative control (IgG). Protein was not incubated with antibodies, as a positive control (Input); Protein was also incubated with anti-Huwe1 antibody (a) or anti-Gadd45b antibody (b), followed by western blot. Note that the interaction of Huwe1 and Gadd45b at 24 h after reperfusion compared to normal neuron cells. c Co-treatment with CHX and lentivirus-GFP-scramble stably decreased the expression of Gadd45b since 0–60 min at 24 h after reperfusion. Conversely, co-treatment with CHX and shRNA-Huwe1 stably increased the expression of Gadd45b since 0–60 min. Tubulin was used as an internal control for western blot; &p < 0.05 versus LV-GFP-ctrl at different time point for CHX