Figure 4.
RAS Regulates PD-L1 Expression through TTP
(A) qPCR analysis of H358 cells following siRNA-mediated knock-down of TTP (24 hr) followed by MEK inhibition (24 hr). Mean ± SEM of two independent experiments.
(B) qPCR analysis of ER-KRASG12V type II pneumocytes treated for 24 hr in starvation medium. Mean ± SEM of three independent experiments.
(C) qPCR analysis of RNA-IP immunoprecipitates from H358 cells. Mean ± SEM from biological triplicates.
(D) Western blotting analysis of H358 cells expressing the indicated constructs. 6.5 hr post-transfection, cells were treated with DMSO or MEK inhibitor for an additional 16 hr. Arrow indicates Myc-TTP. Data are representative of two independent experiments.
(E) Western blotting analysis of immunoprecipitations from H358 cells transfected with Myc-TTP. 6.5 hr post-transfection, cells were treated with DMSO or MEK inhibitor for an additional 16 hr. Arrow indicates Myc-TTP; ∗ indicates co-precipitating protein. Data are representative of two independent experiments.
(F) qPCR analysis of TTP WT or TTP KO MEFs treated with okadaic acid or DMSO for 2 hr. Mean ± SEM of two independent experiments.
Abbreviations and quantities: EtOH, ethanol vehicle; 4-OHT, 100 nM; okadaic acid, OA, 1 μM; MEK inhibitor, GSK1120212, 25 nM. ∗∗∗∗p < 0.0001, ∗∗∗p < 0.001, ∗∗p < 0.01. Unpaired, two-tailed Student’s t tests. See also Figure S4.