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. 2015 Dec 18;5:18462. doi: 10.1038/srep18462

Figure 7. Overexpression of mutant BAP1 upregulates Hsp response and increases transactivity of BAP1.

Figure 7

(a) Immunoblot analysis demonstrated increase of Hsp90 and Hsp70 at cellular level due to aggregation. GAPDH was used as a loading control. (b) I47F, F81V, G178V induced significantly higher level of Hsp90 as compared to wild type (WT). Substantial increase of Hsp70 was observed in aggregating mutants A95D and G178V. (c) Real time PCR analysis determined up-regulation of Hsp70 and Hsp90 at gene level. Values were normalized to GAPDH. Both I47F and A95D showed higher level of Hsp90 gene expression. A95D displayed substantial increase in Hsp70. Representative results are of four independent experiments. EV represents empty vector transfected cells. ***p < 0.001, **p < 0.01 and *p < 0.05, WT. (d) Q PCR analysis showed transactivation of MCM3, TP53I3 and CDKN1B gene expression in mutants I47F, F81V and A95D as compared to wild type (WT) in HEK293T transfected cells. Wild type BAP1 normally acts as a repressor for these genes. Values were normalized to GAPDH. Representative results of three independent experiments °p < 0.1, ***p < 0.001, **p < 0.01 and *p < 0.05, WT.