IL-4Rα shRNA (m) Lentiviral Particles

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	PyVT-R221a cells were isolated from a polyoma middle T oncoprotein (PyMT) tumor as previously described [13]. The R221a line was authenticated by expression of the polyoma antigen, and the morphology and growth rates of both cell lines were consistently monitored. Cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) containing 10% fetal bovine serum (FBS, Atlanta Biologicals) at 37°C with 5% CO2. Control non-target and specific murine IL4Ra-targeted shRNA lentiviral particles (Santa Cruz Biotechnologies) were used to infect R221a cells. Post-infection, shRNA-expressing cells were selected with puromycin dihydrochloride (Sigma). The percent of IL4Rα protein knockdown was calculated by densitometry in Adobe Photoshop® following western blot analysis.

Protocol tips

PyVT-R221a cells were isolated from a polyoma middle T oncoprotein (PyMT) tumor as previously described [13]. The R221a line was authenticated by expression of the polyoma antigen, and the morphology and growth rates of both cell lines were consistently monitored. Cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) containing 10% fetal bovine serum (FBS, Atlanta Biologicals) at 37°C with 5% CO2. Control non-target and specific murine IL4Ra-targeted shRNA lentiviral particles (Santa Cruz Biotechnologies) were used to infect R221a cells. Post-infection, shRNA-expressing cells were selected with puromycin dihydrochloride (Sigma). The percent of IL4Rα protein knockdown was calculated by densitometry in Adobe Photoshop® following western blot analysis.

Publication protocol

PyVT-R221a cells were isolated from a polyoma middle T oncoprotein (PyMT) tumor as previously described [13]. The R221a line was authenticated by expression of the polyoma antigen, and the morphology and growth rates of both cell lines were consistently monitored. Cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) containing 10% fetal bovine serum (FBS, Atlanta Biologicals) at 37°C with 5% CO2. Control non-target and specific murine IL4Ra-targeted shRNA lentiviral particles (Santa Cruz Biotechnologies) were used to infect R221a cells. Post-infection, shRNA-expressing cells were selected with puromycin dihydrochloride (Sigma). The percent of IL4Rα protein knockdown was calculated by densitometry in Adobe Photoshop® following western blot analysis.

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