Skip to main content
. 2021 Sep 8;6(17):e147057. doi: 10.1172/jci.insight.147057

Figure 5. CD36 deacylation induced by lipolysis.

Figure 5

(A) Concentration of FFA in culture medium 5 hours after lipolysis induction in 3T3-L1 adipocytes untreated or treated with an inhibitor of S-acylation, ML211 (30 nM). Data are shown as mean ± SEM; *P < 0.05 (Student’s t test). (B) ABE assay (Supplemental Figure 4A) on 3T3-L1 1adipocytes untreated or induced to undergo lipolysis for indicated time (minutes). Note that IP of S-acylated CD36, specifically observed upon hydroxylamine (HA) treatment, was partly inhibited by lipolysis. Immunoblotting for calnexin and ANX2 from the same extracts indicated constant ANX2 acylation and equal loading. Quantification is on the right. (C) CD36 metabolic labeling (Supplemental Figure 4B). After incubation of live 3T3-L1 adipocytes with alkynylated LCFA analog (0.1 mM 17-ODA, 12 hours), de novo S-acylation of CD36 was detected by IP with anti-CD36 antibodies, subsequent click chemistry with IRDye800-azide probe (IRDye800-N3), and SDS-PAGE. Note that IP of S-acylated CD36, specifically observed upon HA treatment, was inhibited by lipolysis induction (IBMX/forskolin/isoproterenol). Arrow: glycosylated CD36. Arrowhead: nonglycosylated CD36. CD36-IR800 quantification is on the right. (D) Probing of the IP blot from C with PHB and ANX2 antibodies, and subsequently with CD36 antibodies, demonstrated a decrease in PHB and ANX2 association with CD36 concomitant with a decrease of CD36 acylation upon lipolysis induction.