![Changes in p62, UCP1, DRP1, FIS1, LC3B, and mitochondrial condition in PLD2-inhibited or adipocyte Pld2-ablated mice.(A and B) Representative Western blot images (top) and quantification (bottom) of p62, UCP1, and β-Actin, or GAPDH in ingWAT (A) and BAT (B) from vehicle- or CAY10594-injected obese mice (n = 4/group). (C) The area (left) and cristae length (right) of mitochondria in BAT from HFD-fed (12 wk) control and Pld2 Ad-KO mice (n = 6/group). (D) The area (left) and cristae length (right) of mitochondria in BAT from vehicle- or CAY10594-injected obese mice (n = 6/group). (E) Representative seahorse extracellular flux assays measuring OCR in primary brown adipocytes treated with vehicle or CAY10594 (10 µM) for 5 d (n = 3/group). (F) Representative seahorse extracellular flux assays measuring OCR in primary brown adipocytes from control and Pld2 Ad-KO mice (n = 3/group). (G) Representative Western blot images of pDRP1S616, pDRP1S637, DRP1, FIS1, and β-Actin in primary brown adipocytes treated with CAY10594 (10 µM) and carbonyl cyanide m-chlorophenylhydrazone (CCCP; 20 µM) for the indicated time lengths. (H) Representative Western blot images of p62, LC3B, and β-Actin in primary brown adipocytes treated with CAY10594 (10 µM) for the indicated time lengths. The data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-tailed Student’s t test (A [bottom], B [bottom], and C–F). Data are representative of two (G and H) or three independent experiments (A and B). Veh, vehicle.](https://cdn.rupress.org/rup/content_public/journal/jem/219/2/10.1084_jem.20211523/3/jem_20211523_figs3.png?Expires=1767715551&Signature=SqT-WdVNx7XQy-bH8bdEHFtDhqwSPTGuYyyiE4fItDNLhLnfOHPORW1cHaLTv-fknuLLvoqQWT9-Is4BgTfIKXYivzFJ7Twe6~OWs5-isnODwJLELLU9HyqtvI0ZhYgTdNeGetG3EURBEgF5Qnk55qTheJvYt9oCcXdzoxJgpgI1qlfTtQSk6s2HhqMefAY~yvxaePrMlMfhE0tqwO2z9-xIgEoRZDQypjk1UMkttLpEmxD5FnraE3M3bl068PksB-3T-15A1w8O9aTO8atEKUg-fZhRjptxhL7FWi-wIbGScn2x8anHVclFNG8fkHuSP6lfM4rn1oogOFTc-lHNpA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Changes in p62, UCP1, DRP1, FIS1 , LC3B, and mitochondrial condition in PLD2-inhibited or adipocyte Pld2-ablated mice. (A and B) Representative Western blot images (top) and quantification (bottom) of p62, UCP1, and β-Actin, or GAPDH in ingWAT (A) and BAT (B) from vehicle- or CAY10594-injected obese mice (n = 4/group). (C) The area (left) and cristae length (right) of mitochondria in BAT from HFD-fed (12 wk) control and Pld2 Ad-KO mice (n = 6/group). (D) The area (left) and cristae length (right) of mitochondria in BAT from vehicle- or CAY10594-injected obese mice (n = 6/group). (E) Representative seahorse extracellular flux assays measuring OCR in primary brown adipocytes treated with vehicle or CAY10594 (10 µM) for 5 d (n = 3/group). (F) Representative seahorse extracellular flux assays measuring OCR in primary brown adipocytes from control and Pld2 Ad-KO mice (n = 3/group). (G) Representative Western blot images of pDRP1S616, pDRP1S637, DRP1, FIS1, and β-Actin in primary brown adipocytes treated with CAY10594 (10 µM) and carbonyl cyanide m-chlorophenylhydrazone (CCCP; 20 µM) for the indicated time lengths. (H) Representative Western blot images of p62, LC3B, and β-Actin in primary brown adipocytes treated with CAY10594 (10 µM) for the indicated time lengths. The data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-tailed Student’s t test (A [bottom], B [bottom], and C–F). Data are representative of two (G and H) or three independent experiments (A and B). Veh, vehicle.