Impact of STAU1 depletion on the protein expression profile of rhabdomyosarcoma cell lines

Two human rhabdomyosarcoma cell lines (RD, RH36, RH30, and RH18) and one none-transformed skeletal muscle cell line (HSMM-C3) cell line were studied for the impact of Staufen1 depletion on their protein expression profile. Proteomics was performed for control and Staufen1 knockdown cells (n=3). For this, we generated STAU1 knockdown cell lines using a mixture of three independent STAU1-targeting lentiviral-shRNA constructs. cells were cultured for 72 hours and then protein extraction was performed as follow: Cells were lysed in 1X RIPA buffer (20 mM Tris-HCl (pH 7.5), 150mM NaCl, 1mM Na2- EDTA, 1mM EGTA, 1% NP-40, 1% sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1mM β-glycerophosphate, 1mM Na3 VO4, 1µg/ml leupeptin). Protein concentration was tested according to the BCA assay protocol (Thermofisher Scientific) and samples were used in proteomics experiment.