Fusion fx

Protein isolation was performed by lysis of the cells in RIPA buffer (Sigma-Aldrich, St. Louis, MO, USA), supplemented with complete protease and phosphatase inhibitors (Roche, Basel, Switzerland). Proteins were resolved on a 7.5% Bis-Tris gel and blotted onto a 0.45 µm nitrocellulose membrane (GE Healthcare Life Sciences, Chalfont St. Giles, UK). After blocking the membranes with 5% skimmed milk in Tris-buffered Saline Tween (TBS-T), they were incubated overnight with the respective primary antibodies (Supplementary Table S1). Membranes were washed 3× for 10 min with TBS-T. Secondary antibody incubation was performed for 1 h at RT, and membranes were subsequently washed 3× for 10 min with TBS-T. Amersham ECL Prime Western Blotting Detection Reagent was used for the chemiluminescent detection (GE Healthcare Life Sciences) and captured with the imaging device Fusion FX.

Protein isolation was performed via lysis of the cells in RIPA buffer (Sigma-Aldrich, St. Louis, MO, USA) supplemented with complete protease and phosphatase inhibitors (Roche, Basel, Switzerland). The proteins were separated on a 7.5% Bis-Tris gel and blotted onto a 0.45 µm nitrocellulose membrane (GE Healthcare Life Sciences, Chalfont St. Giles, UK). After blocking the membranes with 5% skimmed milk in Tris-buffered saline tween (TBS-T), they were incubated overnight with the respective primary antibodies (Supplementary Table S1). The membranes were washed 3× for 10 min with TBS-T. Secondary antibody incubation was performed for 1 h at RT and the membranes were subsequently washed 3× for 10 min with TBS-T. Amersham ECL Prime Western Blotting Detection Reagent was used for the chemiluminescent detection (GE Healthcare Life Sciences) and captured with the imaging device Fusion FX.