Figure 3.
Effect of mutations on STAT3 homodimerization assessed using BRET reporter constructs. (A) STAT3 WT and mutant BRET constructs were generated that contained (NLuc inserted at the N-terminus of STAT3 and a FlAsH motif inserted within a flexible loop in the DBD immediately after residue 425 (NSF). A control STAT3 WT construct (NS) was generated that contained the NLuc but lacked the FlAsH motif. (B) A schematic of the mechanism for an increased BRET signal following STAT3 Y705 phosphorylation and homodimerization of an NSF construct is shown. (C) MEF−/− cells transiently transfected with the indicated NSF construct or control WT construct (NS) were incubated without or with IL6/sIL6R and used to measure intracellular mBU in real time. The mean ± SEM of 4 assays is shown (∗P < .0001; Student t test). mBU, milli-BRET units.

Effect of mutations on STAT3 homodimerization assessed using BRET reporter constructs. (A) STAT3 WT and mutant BRET constructs were generated that contained (NLuc inserted at the N-terminus of STAT3 and a FlAsH motif inserted within a flexible loop in the DBD immediately after residue 425 (NSF). A control STAT3 WT construct (NS) was generated that contained the NLuc but lacked the FlAsH motif. (B) A schematic of the mechanism for an increased BRET signal following STAT3 Y705 phosphorylation and homodimerization of an NSF construct is shown. (C) MEF−/− cells transiently transfected with the indicated NSF construct or control WT construct (NS) were incubated without or with IL6/sIL6R and used to measure intracellular mBU in real time. The mean ± SEM of 4 assays is shown (∗P < .0001; Student t test). mBU, milli-BRET units.

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