Image lab software version 4.0.1 build 6

Cells were lysed in SDS (sodium-dodecyl sulfate) lysis buffer (Szymanska, Fosdahl et al. 2016) supplemented with 10 μg/ml protease inhibitor and phosphatase inhibitor cocktails and subjected to homogenization using a QIAshredder column (QIAGEN, Valencia, CA, USA). Sample buffer (Szymanska, Fosdahl et al. 2016 ) was added to the lysates before incubation for 10 min at 95°C. Proteins were separated by SDS gel electrophoresis and protein blotting using 10% Mini-PROTEAN®TGX TM Precast Gels and Trans-Blot® Turbo TM Transfer System (Bio-Rad Laboratories, Hercules, CA, USA). The membranes were incubated with primary and secondary antibodies diluted in 1% Blotting-Grade Blocker (Bio-Rad Laboratories) for 2 hours at room temperature or over night at 4°C before detection of proteins using Super Signal West Dura Extended Duration Substrate from Thermo Fisher Scientific (Waltham, MA USA) and visualization using the ChemiDoc MP System (Bio-Rad Laboratories). Image Lab Software version 4.0.1 build 6 (Bio-Rad Laboratories) and Adobe Photoshop (Adobe Systems, San José, CA, USA) were used for image processing.

Cells were lysed in SDS (sodium-dodecyl sulfate) lysis buffer (Szymanska, Fosdahl et al. 2016 ) containing 10 μg/ml 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF). Lysates were subjected to homogenization using a QIAshredder column (QIAGEN, Valencia, CA, USA). Sample buffer (Szymanska, Fosdahl et al. 2016 ) was added to the lysates before incubation for 10 min at 95°C. Proteins were separated by SDS-gel electrophoresis using 10% Mini-PROTEAN®TGXTM Precast Gels from Bio-Rad Laboratories (Hercules, CA, USA). Afterwards, proteins were blotted onto a nitrocellulose membrane using Trans-Blot® TurboTM Transfer System (Bio-Rad Laboratories). The membranes were incubated with 5% Blotting-Grade Blocker in TBS-Tween 20 (Bio-Rad Laboratories) before incubation with primary and secondary antibodies diluted in 1% Blotting-Grade Blocker in TBS-Tween 20 for 2 hours at room temperature or over night at 4°C. For detection of proteins, Super Signal West Dura Extended Duration Substrate from Thermo Fisher Scientific was used. Visualization was achieved with the ChemiDoc MP System (Bio-Rad Laboratories). Image Lab Software version 4.0.1 build 6 (Bio-Rad Laboratories) and Adobe Photoshop (Adobe Systems, San José, CA, USA) were used for image processing.