Skip to main content
. Author manuscript; available in PMC: 2020 Jul 6.
Published in final edited form as: FASEB J. 2020 Apr 15;34(6):8265–8282. doi: 10.1096/fj.201903100RR

FIGURE 4.

FIGURE 4

Deficiency of DRP1 causes abnormal retention of nascent micro-LDs in ER in adipose tissue. A, Co-IF staining with anti-Perilipin-3 (PLIN3, green) and anti-PLIN1 (red) antibodies in the BAT from Adipo-Drp1flx/flx mice and their littermate controls after cold exposure. Yellow signals in the merged panel indicate merging of the newly formed LDs with the preexisting LDs in the cytosol. B, Pearson’s correlation coefficient (PCC) for colocalization of PLIN3 and PLIN1 in (A) (Representative of three trials is shown. The PCC of 5 fields were analyzed and presented as mean ± SEM, Student’s t test, *P < .05). C, Western blotting analysis of PLIN2 and KDEL protein levels in ER fraction isolated from the BAT of Adipo-Drp1flx/flx mice and their littermate controls after cold exposure. D, Quantification of relative band intensity for PLIN2 (normalized to KDEL) in (C) (n = 3 per group for each experiment, representative of three trials is shown. The data are represented as mean ± SEM, Student’s t test, *P < .05). E, Triacylglycerol (TAG) levels in the ER fraction of the adipose tissues from Adipo-Drp1flx/flx mice and their littermate controls. TAG levels were normalized to protein concentration (n = 3 per group for each experiment, which were performed in triplicate, data are represented as mean ± SEM, Student’s t test, ***P < .001). F, Co-staining of BODIPY-C12 (red) and ER-tracker (green) in primary adipocytes differentiated from SVF of Adipo-Drp1flx/flx mice and their littermate controls. Cells were infected with lentivirus expressing GFP or DRP1-GFP during differentiation. The cells were stained with BODIPY-C12 for 10 minutes and chased in BODIPY-C12 free medium for 1 hour. The cells were stained with ER-tracker during the 1 hour chase phase. G, Pearson’s correlation coefficient (PCC) for the colocalization of BODIPY-C12 and ER-tracker in (F) (Representative of three trials are shown. The PCC of 5 fields were analyzed and represented as mean ± SEM, One-way analysis of variance (ANOVA), *P < .05, **P < .01, no significant difference between the first and the last group)