Post on 07-Nov-2018
Supplementary Fig. 2. Regulation of mitochondrial biogenesis and representative
photomicrographs of confocal microscopy detection for double immunofluorescence of UCP1
and VEGF-A in ingWAT of DJ-1 KO mice. (A) Transcript levels of indicated genes required for
mitochondrial biogenesis (Nrf1, Tfam, CoxIV, and Cox8b) from tissue lysates of ingWAT. (B)
Western blot (left) and densitometry analysis (right) of indicated proteins involved in
mitochondrial electron transport chain (COX-IV, ATP5B and ETFA) in ingWAT extracts. (C)
Representative red MitoTracker staining on ingWAT sections. Nuclei were counterstained with
DAPI (blue). Scale bar, 50 µm. (D) Immunoblotting (top) and quantitation (bottom) of indicated
proteins implicated in mitochondrial fusion (Mfn1) and fission (Drp1) in ingWAT tissue lysates.
(E) Double immunofluorescence detection with antibodies against brown adipocyte marker
UCP1 (green) and VEGF-A (red) on representative sections of ingWAT. Nuclei were stained
with DAPI (blue). Co-localization is shown by merged images. Scale bar, 50 µm. Results are
representative of at least three independent experiments. Graphical data show densitometry
analyses normalized to actin (loading control). Gene expression levels were normalized to β-
actin. Data are presented as the means ± S.D. Data were analyzed by using ANOVA Tukey’s
multiple comparisons or Student’s t-test (if two groups). *p < 0.05, **p < 0.01 compared to WT
counterpart (A, B, D); †p < 0.05, ††p < 0.01 compared to WT-ND group (A, B, D).