Extended Data Fig. 12: CREM KO does not impact IL-15 receptor (IL-15R) proximal signaling but enhances the metabolic fitness of CAR70/IL-15 NK cells.
From: CREM is a regulatory checkpoint of CAR and IL-15 signalling in NK cells
(a) Whole cell lysates from NK cells that were either wild-type (WT) or knockout (KO) for CREM 0, 30 min and 2 h following stimulation with IL-15 (5000 pg/ml) as analyzed by western blot for IL-15R proximal signaling components. β-actin serves as a loading control (n = 5 donors); (b) Densitometry analysis quantifying the relative band intensity of pSTAT3 and pSTAT5 normalized to total STAT3 and STAT5, respectively (n = 5 donors); (c) CISH expression in NK cells that were either WT or CREM KO that were either unstimulated or stimulated with IL-15, assessed by qPCR (n = 3 donors); (d-f) Measures of extracellular acidification rate (ECAR) upon addition of glucose, oligomycin (oligo), and 2-deoxy-D-glucose (2-DG) (d) and quantified basal glycolysis (e) and glycolytic capacity (f) of NK cells in the indicated groups (n = 3 donors); (g-i) Measures of oxygen consumption rate (OCR) upon addition of oligo, FCCP, and rotenone and antimycin A (R/A) (g) and quantified basal respiration (h) and maximal respiration (i) of NK cells in the various conditions (n = 3 donors). ns: non-significant. Statistical comparisons were performed using two-way ANOVA with Šídák correction (b,c), and one-way ANOVA with Tukey correction (e,f,h,i). Data are represented as mean ± SEM.
