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. 2005 Sep 15;102(39):14080–14085. doi: 10.1073/pnas.0500099102

Fig. 4.

Fig. 4.

Phosphorylation of Rb on Ser-795 is diminished by DNCdk4 expression. (a) Time course of phosphorylation of Rb on Ser-795 and effects of DNCdk4 expression. Animals were injected unilaterally with DNCdk4-expressing virus. At the indicated times after 4VO, ipsilateral (with virus) or contralateral (no virus) nuclear hippocampi proteins were extracted and analyzed for Ser-795 phosphorylation by Western blot. Total Rb was used as loading control. (b and c) Comparison of the effects of DNCdk2/4/5 and GFP on Ser-795 Rb phosphorylation 12 h after ischemia. Positive control refers to nuclear hippocampal proteins extracted from noninjected animal O(-Ser) and 12 h (+Ser) after reperfusion. All lanes in c are from the same Western blot; Coomassie blue staining as loading control. (d) Time course of phosphorylation of Rb on Ser-795 in vitro. CGNs were subjected to 16 h of hypoxia, in the presence of MK801, followed by up to 12 h of reoxygenation; actin was used as loading control. (e) Expression of mutant Rb provides significant protection from hypoxia-induced delayed neuronal death in vitro. Quantitation of survival after 16–18 h of hypoxia followed by 24-h reoxygenation, in the presence of MK801. The data are mean ± SEM (n = 3). * denotes significance (P < 0.05, t test hypoxia GFP vs. hypoxia mut.Rb)