Skip to main content
. 2019 Jan 15;11(1):97. doi: 10.3390/cancers11010097

Figure 5.

Figure 5

Searching for the E3-ligase that mediates degradation of BRCA2 upon hyperthermia. (A) Effect of hyperthermia on BRCA2 levels upon knock-down of the Heat-shock Protein (HSP)-associated E3-ligase, C terminus of HSC70-Interacting Protein (CHIP), in U2OS cells using two different siRNAs. Arrow on the right next to the CHIP panel indicates the predicted position of the CHIP protein. (B) Effect of hyperthermia on BRCA2 levels upon knock-down of the SUMO-E3 ligases Protein Inhibitor Of Activated STAT( PIAS) 1-4 in U2OS cells using siRNAs. (C) Effect of hyperthermia on BRCA2 levels upon siRNA knock-down of DNA-repair-related, SUMO-targeted ubiquitin ligases, Ring Finger (RNF) 4 and RNF111. (D) Detection of BRCA2 and BRCA1 in UWB1.289 (BRCA1-null) cells with or without complementation of wild type BRCA1 (wtBRCA1). (E) Detection of BRCA2 in U2OS cells treated with increased doses of the neddylation-inhibitor MLN4924, added 30 min prior to hyperthermia. (F) Detection of BRCA2 in U2OS cells treated with or without hyperthermia in the presence of proteasome inhibitor MG132 (10 µM) or the E1-activating enzyme inhibitor PYR-41 (10 µM). Inhibitors were added 1 h prior to hyperthermia treatment, and left on for the duration of the treatment.