Western blotting was performed using the Invitrogen Novex Mini-Cell system with NuPAGE 4 to 12% bis-tris gels. Proteins were transferred to nitrocellulose membranes (0.45 μm; Bio-Rad) and blocked using LI-COR blocking buffer at a 1:1 ratio with PBS for 1 hour at room temperature. Primary antibodies were mixed in tris-buffered saline (TBS)–0.1% Tween 20 (TBST) with 2% BSA and incubated with membranes overnight at 4°C on a shaker. Membranes were then washed three times for 10 min in TBST and incubated with fluorophore-conjugated secondary antibodies in TBST with 2% BSA for 1 hour at room temperature. Membranes were washed three times for 10 min in TBST and imaged using a LI-COR Odyssey imaging system. Quantification of bands was done using ImageStudio software (LI-COR).

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