Western Blot Analysis Protocol

Immunoblot analysis was performed as described previously [8 (link)]. In short, all lysates for immunoblot analysis were prepared by lysing cells in PBS‐1%TritonX‐100 buffer supplemented with Protease Inhibitor Cocktail and Phosphatase inhibitor (Roche, Basel, Switzerland). After that, the resulting suspension was incubated on ice for 10 min, centrifuged, and the supernatant collected. Tissue lysates were made in the mentioned lysis buffer using Precellys 24 tissue homogenizer (Bertin instruments) and the Precellys kit (#KT03961‐1‐003.2). Proteins were separated on gradient SDS gels (BioRad) and transferred to nitrocellulose membranes (BioRad #1704158). Transfer quality was evaluated with Ponceau staining (Sigma, #P7170). Immunoblots were quantitated using image j (Bethesda, MD, USA). Statistical significance was determined via a one‐sided t‐test, unless specified otherwise in the figure legends.