Cell lysates containing a quarter of a volume of 4X NuPAGE LDS sample buffer (NP0007) supplemented with 5% βmercaptoethanol, were heated at 95°C for 5 minutes. 15 to 20 μg of samples were loaded onto pre-cast 4-12% Bis-Tris midi 20W or 26W gels (Thermofisher Scientific, Cat# WG1402BOX or WG1403BOX) and resolved at 130 V for 2 hours with NuPAGE MOPS SDS running buffer (Thermofisher Scientific Cat# NP0001-02). Proteins were electrophoretically transferred onto a 0.45 µm nitrocellulose membrane (GE Healthcare, Amersham Protran Supported 0.45 mm NC) at 90 V for 90 min on ice in transfer buffer (48 mM Tris base and 39 mM glycine supplemented with 20% methanol). The transferred membrane was blocked with 5% (w/v) skim milk powder dissolved in tris-buffered saline with tween (TBS-T) (50 mM Tris base, 150 mM sodium chloride (NaCl), 0.1% (v/v) Tween-20) at room temperature for 1 hour. Membranes were washed three times with TBS-T and were incubated in primary antibody overnight at 4°C. Prior to secondary antibody incubation, membranes were washed three times for 15 minutes with TBS-T. The membranes were incubated with secondary antibody for one hour at room temperature, protected from light. Thereafter, the membranes were washed with TBS-T three times with a 15-minute incubation for each wash, and protein bands were acquired via near-infrared fluorescent detection using the Odyssey CLx imaging system and quantified using Image Studio software. Graphs were generated using Graphpad Prism version 8 software.