As n Chstes we examined the effects of ectopic expression of bcl-2 and bcl-2 in weight BH4 Dom ne of HIF-1a protein in post-transcriptional level, deleted with the proteasome inhibitor MG132 gel. The accumulation of HIF 1a protein of MG132 treatment was identical in all the cells independently Ngig the status of the bcl 2, indicating that neither the shape nor affect transferred wt bcl R788 Fostamatinib 2 protein synthesis HIF 1a. be directly tested whether bcl 2 in hypoxic cells condition with modulation of HIF Proteinstabilit t is assigned 1a ma s we. the half-life of HIF 1a protein In Figures 5c and d show a decrease in the time h hangs from the HIF 1a in cells after treatment with the inhibitor of protein synthesis, cycloheximide after hypoxia was observed.
Densitometric analysis revealed that, as previously indicated, 21 bcl 2 overexpression the duration of HIF 1a protein half 155-455 min obtained Ht under hypoxic conditions. In contrast, no significant difference in the lives BMS-790052 of a half was HIF 1a protein between the embroidered and the stable clones bcl 2 of its BH4 Dom ne gel Observed deleted. And a reduction of the ubiquitination in HIF 1a transfectants bcl weight 2, as compared to control cells was observed, the modulation frequency of the HIF 1a Hnlicher ubiquitination transfectants carrying deletion of the BH4 Dom ne and control cells was observed. Independently, the effect of two mutations on bcl HIF 1/VEGF signaling Dependent.
Of their relevance in apoptosis The M opportunity That the effect of two of bcl exclude his 1/VEGF expression and activation of HIF k Nnte consequence of a different modulation of the apoptotic pathway by ectopic expression of various forms of bcl 2 S, we evaluated the response of the cells to camptothecin, a drug capable of the apoptotic program activated in several experimental models, 22. CPT treatment induces apoptosis in about 50% of non-transfected cells or cells transfected with the empty vector, w While it overexpresses a very small percentage of apoptosis in the cells of Bcl 2 weight induced. CPT-induced apoptosis was also in cells encoding with expression vectors BH1, BH2 and BH4 Dom ne transfected observed by Bcl-deletion mutant 2, bcl two point mutants of highly conserved residues within the BH1 BH2 or areas or dicodon mutants in the BH4 Dom-ne.
On the other hand, the rate of apoptotic Vorg length In cells with Bcl two mutations in residues July 6 10 11 16 and 17 of the BH4 Dom ne was to that much Similar to CPT treatment observed in cells overexpressing bcl wt transfected 2, the best firmed that these residues are not necessary for the anti-apoptotic Bcl second 6,7,23 These results were also obtained by cleavage of poly polymerase after treatment with CPT in cells bcl 2 BH4 Dom ne gel Deleted best Entitled, but not in cells overexpressing bcl weight second Zus Tzlich was cleavage of PARP in cells overexpressing bcl significantly reduced two mutated amino Acids July 6, 10 11 and 16 17 ne in the BH4-Dom. As n Chstes we examined whether the two bcl-induced cell death protein and the production of reactive oxygen species mutated in hypoxia. Foreign non-exposure of cells to hypoxia for 24 hours Sen either apoptosis or ROS generation or ma Or transmitted in cells overexpressing bcl weight or 2. In all cells were o10% of apoptosis and ROS observed o5% after 24 h hypoxia against