Figure 1
From: Sec-containing TrxR1 is essential for self-sufficiency of cells by control of glucose-derived H2O2
Characterization of expression levels, Sec incorporation and total cellular enzyme activity of TrxR in MEFs with depleted or reconstituted Txnrd1 variants status. (a) Protein expression levels of TrxR1 incubated with or without 25 nM selenite supplementation in the medium for 24 h were analyzed by immunoblotting using reducing SDS-PAGE (top panel). Unspecific bands are indicated by asterisks (*) and TrxR1 dimeric bands are indicated by an arrow in parentheses. Endogenous (‘TrxR1’) and reconstituted (‘SF-TrxR1’) variants are indicated between the 55 and 70 kDa weight markers. Ponceau S staining was used as loading control (bottom panel). (b) Sec incorporation was determined using autoradiography of 75Se-labeled selenoproteins. The total proteins of lysed cells were analyzed on a reducing SDS-PAGE gel and exposed to a phosphor screen (top panel). Coomassie staining was used as loading control (bottom panel). (c) Total cellular TrxR activity was determined using a specific Trx-linked insulin disulfide reduction assay, with proteins of the same cell lysates as shown in (a) (n=2)
