Trxr1

Pancreatic tail tissues were fixed in 4% paraformaldehyde, paraffin-embedded, and sectioned. Paraffin sections were conventionally dewaxed in xylene, rehydrated through decreasing concentrations of ethanol, and washed in phosphate buffered saline, followed by permeabilization with 1% Triton X-100. After blocking in 3% BSA for 45 minutes, the sections were incubated overnight in the insulin (mouse monoclonal, 1:400; Proteintech). Then, the sections were treated with FITC-conjugated Affinipure goat anti-mouse (1:200; Huaxingbochuang) for 1 hour before they were incubated overnight in NOX1 (1:50; Proteintech), NOX2 (1:50; Proteintech), NOX4 (1:50; Proteintech), TrxR 1 (Trx1) (1:50; Proteintech), TrxR1 (1:200; Cell Signaling Technology), Trx2 (1:50; Proteintech), TrxR2 (1:50; Proteintech), or TXNIP (1:50; Proteintech) and treated with TRITC-conjugated Affinipure goat anti-rabbit (1:200; Huaxingbochuang) for 1 hour. Antifluorescence quenching mounting medium with DAPI was mounted on the sections. Images were viewed using the Leica SP8 confocal microscope.