Fig. 6: Axl KO and iFAXLKO mouse models are protected from diet-induced obesity (DIO).

A Transgenic cassette and subsequent global deletion of AXL receptor (upper) and representative western blot of three independent experiments of AXL receptor in tissues of global Axl KO (homozygous) mice (lower). B Weekly body weight curves of Axl KO homozygous (AXLKO Hom), Axl KO heterozygous (AXLKO Het) and WT female littermates on HFD challenge. HFD was initiated at 8 weeks of age, mice were acclimatized to room-temperature conditions (n = 27, 29, 31; respectively). *p < 0.05, ** p < 0.01, ***p < 0.001 and ****p < 0.0001 (for WT vs. AXLKO Hom). ^& p < 0.05, ^&&p < 0.01, ^&&& p < 0.001 and ^&&&& p < 0.0001 (for AXLKO Het vs. AXLKO Hom), two-way ANOVA was performed. C Representative H&E staining images iBAT depots of Axl KO and WT littermates after 24 weeks of HFD. iBAT depots of all Axl KO and WT mice were imaged. D Measurements of inguinal WAT (ingWAT), gonadal WAT, and iBAT fat depot weights (after 24 weeks of HFD), lean mass (via EchoMRI – after 9 weeks of HFD), and food consumption measurements of Axl KO and WT littermate controls, after 24 weeks of HFD challenge (WT n = 8; Axl KO n = 7) (E) VO2 metabolic cage measurements in Axl KO and WT female mice 24 weeks after HFD challenge (n = 6/group). F Body weight, total cholesterol, free fatty acids (FFA), fasting blood glucose, iBAT weight, and liver weight of Axl KO and WT littermate controls, after 32 weeks of a high-fructose high-fat (HF/HF) diet challenge that was used to generate a diet-induced obesity and fatty liver accumulation mouse model (WT-chow n = 4; Axl KO-chow n = 6; WT-HF/HF n = 5; Axl KO-HF/HF n = 6). G Weekly weight gain of Axl KO and WT littermate controls, in response to 16-week HFD challenge at room temperature (RT) (n = 25 for Axl KO and n = 30 for WT). H Weekly weight gain of Axl KO and WT littermate controls, in response to 16-week HFD challenge, at thermoneutrality (TN) (n = 10/group). I Body weight of Axl KO and WT littermate controls after treatment with the pharmacological AXL receptor inhibitor BMS-777607 (BMS) or DMSO, in response to HFD challenge. J RNA-Seq analysis of iBAT depots from Axl KO and WT male mice after 25 weeks on HFD and subsequent enrichment analysis of the BATLAS signature score of the differentially expressed genes in Axl KO vs. WT mice (n = 5/group) K) Western blot of the mature fraction of ingWAT depots from control (Axl-floxed Cre-) or iFAXLKO (Axl-floxed CreERT2 + ) mice of AXL receptor in adipocytes. L Weight gain curves of iFAXLKO and littermate control (lox-AXL-lox Cre-) female mice in response to HFD challenge (n = 11/group). M Weight of ingWAT, gWAT, iBAT, and liver in iFAXLKO and floxed-AXL-Cre- control littermates, after 20 weeks of HFD (n = 11/group). N) Glucose tolerance test (glucose dose: 2 g/kg) and O) insulin tolerance test (insulin dose: 1IU/kg) in iFAXLKO and loxP-AXL-Cre- control littermates, after 20 weeks of HFD (n = 11/group). P VO2 metabolic cage measurements in iFAXLKO and littermate control (loxP-AXL (Cre-)) female mice 8 weeks after HFD (n = 6/group). For all graphs, results are presented as average ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. For two-group comparisons (D, M), unpaired two-tailed t-test was performed. For three or more group comparisons (F), one-way ANOVA was performed (Tukey test was applied to correct for multiple comparisons), and for time-course datasets (E, G–I, L, N–P) two-way ANOVA was performed. Statistically significant differences are annotated in graphs.