Fig 2: IL-15–induced PDGFRß expression is mediated by PI3K/AKT signaling. (A and B) Primary NK cells were treated with IL-15 (10 ng/mL) for the indicated times. mRNA levels of PDGFRB at different time points (A) (n = 3) or at 1 h (B) (n = 10) were examined by qPCR. (C and D) Primary NK cells were pretreated with ActD (5 µg/mL) (C) or cycloheximide (CHX, 20 µg/mL) (D) for 1 h, washed twice with RPMI-1640, and then treated with IL-15 (10 ng/mL) for 24 h. Dimethyl sulfoxide (DMSO) was used as a control. Expression levels of PDGFRß were examined by flow cytometry (n = 5). Data shown are representative histograms and percentage of inhibition with the following equation: % inhibition = 100 × [1 - (DMSO-inhibitor)/DMSO]. (E) Primary NK cells were pretreated with wortmannin (1 µM), afuresertib (10 µM), TPCA-1 (1 µM), rapamycin (10 µM), torin1 (10 µM), decernotinib (10 µM), C118-9 (10 µM), STAT5-IN-1 (10 µM), AZD6244 (10 µM), or CI-1040 (10 µM) for 1 h, washed twice with RPMI 1640, and then treated with IL-15 (10 ng/mL) for 24 h. DMSO was used as control. Data shown are percent of inhibition (n = 5). The mean value of the inhibitory rate is shown. (F) Luciferase reporter assay shows that p65 activates PDGFRB gene transcription. (G–I) Binding of p65 to the PDGFRB promoter in IL-15–treated NK cells (G) or resting NK cells (H) as determined by ChIP-qPCR or PCR (I) (n = 3). Data represent three independent experiments. Data shown are means ± SD. NS, not significant. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.