Fig 2: CD36 coordinates with Fyn to phosphorylate IP3R1, inducing lysosomal Ca2+ overload and inflammation in preadipocytes. We constructed a lentivirus containing CD36 with a palmitoylation site mutation and infected the 3T3L1 cells with lentivirus (mt∆CD36 OE preadipocytes). (a) Co-IP for CD36 and Fyn in wtCD36 OE preadipocytes and mt∆CD36 OE preadipocytes (n=3). (b) Western blot analysis of IP3R1 and P-IP3R1 (Tyr353) in wtCD36 OE preadipocytes and mt∆CD36 OE preadipocytes (n=3). (c) Cells were incubated for 30 min in the presence of 0.5 μM fluo-4 AM, a Ca2+ indicator dye, to detect total intracellular Ca2+ (n=3). (d) ER Ca2+ release assay. Cells were incubated with fluo-4 AM (0.5 μM) for 30 min. Intracellular Ca2+ was monitored prior to and following exposure to 1 μM thapsigargin (n=3). (e) Representative images showed double immunofluorescence staining for OG-BAPTA-5N and Lysotracker in NC preadipocytes, wtCD36 OE preadipocytes (treated with or without PP2) and mt∆CD36 OE preadipocytes. (f) Western blot analysis of NF-kB and IL-1β in wtCD36 OE preadipocytes and mt∆CD36 OE preadipocytes (n=3). *p<0.05 compared with the NC group. #p<0.05 compared with the wtCD36 OE group.