Cells were washed with cold PBS then lysed with 300µL cold RIPA buffer (Thermo Fisher Scientific) supplemented with 1x of a cocktail of phosphatase and protease inhibitors (Life Technologies).

Cell lysates were transferred into 1.5ml eppendorf tubes and centrifuged for 0h 5m 0s at 3500rpm to remove cell debris.

Following centrifugation, the supernatant was collected and protein concentration were measured using a BCA or Bradford protein assay (Thermo Fisher Scientific).

Equal amounts of protein lysate, 20µg, were used to perform ELISA assays according to manufacturer's instructions.