1. Silencing Socs1 in the LLC line confers increased response to IFNγ in vitro.
Silencing Socs1 in the LLC line confers increased response to IFNγ in vitro. Two separate shRNAs targeting Socs1 (LLC-sh20 and LLC-sh21) and a nontargeting control vector (LLC-NT) were transduced into LLC cells expressing luciferase. LLC cells were then screened for functional and stable knockdown of Socs1 after 10 d of puromycin selection. (A) Immunoblots showing p-STAT1, total STAT1, and β-ACTIN levels of the LLC-NT, LLC-sh20, and LLC-sh21 cell lines ±100 ng/ml IFNγ after 15 min or 48 h in vitro. The cells were treated with ± 10 ng/ml IFNγ for 24 h followed by isolation of RNA and qRT-PCR. (B–G) mRNA levels of (B) Socs1, (C) Ifngr1, and downstream IFNγ response genes (D) Cxcl9, (E) Cxcl10, (F) Cd274, and (G) Ciita are shown as absolute values (SQ values) normalized to the housekeeping gene Actb from full data sets. Statistics compare the LLC-NT line with other cell lines with IFNγ treatment. Error bars represent the mean of the data ± SEM (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < ) after a two-way ANOVA (B–G). Source data are available for this figure.
Bonnie L Bullock et al. LSA 2019;2:e
© 2019 Bullock et al.