FIGURE 3.

TXN stabilizes Txnip protein. A, Txnip and C247S Txnip transgenes transiently expressed in HEK 293TN and protein levels followed over time after cycloheximide (CHX, 40 μm) translational inhibition. Densitometry (Densitom.) levels reflect expression of Txnip or C247S at each time point relative to the untreated state. B, transfected cells were pretreated with MG132 (PI, 10 μm) or dimethyl sulfoxide (DMSO) vehicle (V) for 10 min prior to the addition of CHX and incubated for 4 h prior to cell lysis. C, 293TN cells were transfected with uniform amounts of Txnip or C247S plasmid and increasing amounts of plasmids carrying TXN or the TXN active site cysteine mutants, C32S and C35S. Empty vector (EV) was added to equalize total plasmid transfected per condition. Ratios indicate relative amounts of each plasmid.