Journal: Diabetes

Article Title: Macrophage HIF-2α ameliorates adipose tissue inflammation and insulin resistance in obesity.

doi: 10.2337/db13-1965

Figure Lengend Snippet: Figure 1—HIF-2a is elevated in the ATMs from db/db mice. A and B: Expression pattern of HIF-2a in the adipose tissue of lean db/+ and obese db/db mice; n = 5 each group. Whole-mount immunofluores- cence analyses of the nucleus (blue), perilipin (green), CD11b (yellow), and HIF-2a (red) were performed on the epididymal adipose tissues of db/+ and db/db mice. C: mRNA expression levels of Hif-2a and Hif- 1a in M1-type (CD11c+ and CD2062) or M2-type (CD11c2 and CD206+) ATMs (F4/80+ and CD11b+) isolated from the epididymal adipose tissues of db/db mice; n = 6. Relative mRNA levels of Hif- 2a, Hif-1a, and M1 and M2 markers were determined using quanti- tative RT-PCR. Expression levels were normalized to the level of Cyclophilin mRNA. Data represent mean 6 SD. *P < 0.05; **P < 0.01 M1- vs. M2-like by Student t test. M, macrophage.

Article Snippet: Epididymal adipose tissues were fixed with 1% paraformaldehyde and blocked with 5% goat serum in PBS Tween for 1 h. Whole-mounted epididymal adipose tissue was incubated with primary antibodies against HIF-2a (1:1,000; Novus, CO), perilipin (1:1,000; Fitzgerald, MA), and CD11b (1:1,000; eBioscience) overnight at 4°C.

Techniques: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction

Journal: Chinese Medicine

Article Title: Unraveling the potential mechanisms of Xiaochaihu decoction alleviates metabolic associated fatty liver disease (MAFLD) by integrated transcriptomics, metabolomics, and network pharmacology

doi: 10.1186/s13020-025-01310-y

Figure Lengend Snippet: XCHD regulates HFSW-induced variations of gene expression in the liver. RNA sequencing analysis was performed ( n = 3). A Volcano map of differentially expressed genes (DEGs). B Biological process (BP), cell component (CC), molecular function (MF) in GO enrichment of DEGs. C KEGG analysis of DEGs. D Pearson correlation analysis between PLINs and significantly differentially expressed lipids and lipid-like molecules. E The protein expression in liver tissue of PLIN2, PLIN3, and PLIN5 was determined by Western blot analysis. F The ratios of PLIN2/β-actin, PLIN3/β-actin, and PLIN5/β-actin are calculated ( n = 3). Data are represented using at least three independent experiments as the mean ± SD. Data are represented using at least three independent experiments as the mean ± SEM. *p < 0.05, **p < 0.01, vs. MOD group

Article Snippet: Antibodies Plin3 (10694–1-AP), Plin5 (26951–1-AP), and β-actin (81115-1-RR) were purchased from Proteintech Group (Rosemont, IL, USA).

Techniques: Gene Expression, RNA Sequencing, Expressing, Western Blot

Journal: The FASEB Journal

Article Title: Perivascular adipose tissue–derived extracellular vesicle miR‐221‐3p mediates vascular remodeling

doi: 10.1096/fj.201901548r

Figure Lengend Snippet: Figure 2. Adipose tissue secretes EVs containing miRNAs. A) Immunostaining for the EV marker CD63 (green) and the adipocyte marker PLIN (red). The white arrows indicate extracellular CD63 stainings by confocal imaging. Scale bar, 15 mm. B) Electron microscopy analysis of EVs secreted by MAT explants, showing a size of ;50–150 nm in diameter. The inset shows higher magnification of EVs. Scale bars, 100 nm. C) EV-related protein markers ALIX (97 kDa), CD63 (50 kDa), TSG101 (44 kDa), and CD9 (25 kDa) as well as adipocyte marker PLIN-A/B (57/46 kDa) were detected by Western blot analysis in MAT-EVs or supernatant CM. Blots are representative of 3 independent experiments. D) Uptake of EVs: Isolated MAT-EVs were labeled with PKH26 (red) and incubated (50 mg/ml) with primary VSMCs for 18 h. Middle, PKH26-labeled MAT-EVs. Nonlabeled MAT-EVs and PKH26 dye (1 3 1026 M) alone were used as controls. Scale bar, 25 mm. E) Total RNA from MAT (cellular) or MAT-EVs was analyzed with a bioanalyzer. Both gels and electropherograms are shown. Y axis of the electropherogram is arbitrary fluorescence unit intensity, and x axis is migration time in seconds.

Article Snippet: Blots were probed with rabbit anti-ALIX (ab186429; Abcam, Cambridge, MA, USA), rabbit anti-CD63 (ab216130; Abcam), rabbit anti-CD9 (ab92726; Abcam), rabbit anti-TSG101 (ab125011; Abcam), mouse anti-CD31 (ab24590; Abcam), mouse anti-CD68 (ab31630; Abcam), and mouse monoclonal antibody for PLIN (10R-2478; Fitzgerald Industries International, Acton, MA, USA) followed by washing and incubationwith the respective secondary antibodies.

Techniques: Immunostaining, Marker, Imaging, Electron Microscopy, Western Blot, Isolation, Labeling, Incubation, Migration

Journal: eLife

Article Title: Arf1/COPI machinery acts directly on lipid droplets and enables their connection to the ER for protein targeting

doi: 10.7554/elife.01607

Figure Lengend Snippet: Figure 3. The COPI machinery localizes to the LD surface. (A) The endogenous COPI machinery stained with αCOP or garz antibodies (red) localizes to LDs in S2 cells. Frequencies of colocalization of αCOP and garz spots with LDs from experiments are higher than expected from a random distribution. (B) The endogenous COPI machinery localizes to LDs in NRK cells. NRK cells stained for βCOP or GBF1 by immunofluorescence (red) show partial colocalization with LDs stained with BODIPY (green). Colocalization of βCOP with LDs in NRK cells is not random. Relative frequencies of βCOP, KDEL receptor and clathrin spots colocalizing with LDs determined in experiments are respectively compared to the frequencies of colocalization from a binomial random distribution. From the two frequencies (experiment vs simulation), a significant overrepresentation of βCOP on LDs is observed, whereas clathrin and KDEL receptor (KDELR) are not found on LDs. For (A) and (B) scale bars are 10 μm (overview) or 1 μm (first inlay) or 250 nm (second inlay). Statistical significance was tested by a student t test with p<0.01 (n = 30). (C) Localization of β’COP (green) to the LD surface (perilipin3, red) using confocal (upper panel) and super-resolution STED microscopy (lower panel). Scale bar = 500 nm (overview) or 100 nm (inlay). (D) Localization of β’COP to LDs is efficiently blocked by treatment of cells with the Arf1 GEF inhibitors brefeldin A or golgicide A. Scale bar = 10 μm (overview) or 1 μm (inlay). DOI: 10.7554/eLife.01607.008 The following figure supplements are available for figure 3:

Article Snippet: Rabbit polyclonal antibodies used: anti-GPAT4 (Wilfling et al., 2013), anti-CCT1 (Wilfling et al., 2013), anti-GBF1 (BD Biosciences, San Jose, CA), anti-KDEL-receptor (KDELR; gift from Dr JE Rothman; Yale University), anti-βCOP (gift from Dr JE Rothman; Yale University), anti-perilipin3 (TIP47; Novus Biologicals, Littleton, CO), anti-αCOP (Abcam, Cambridge, MA), anti-GRP78/BiP (ET-21) (Sigma–Aldrich, St. Louis, MO) and anti-garz (Wang et al., 2012) (gift from Dr A Paululat; University of Osnabrück).

Techniques: Staining, Immunofluorescence, Microscopy

Journal: Obesity (Silver Spring, Md.)

Article Title: Intramyocellular Lipid Droplet Size Rather Than Total Lipid Content Is Related to Insulin Sensitivity After 8-Weeks of Overfeeding

doi: 10.1002/oby.21980

Figure Lengend Snippet: A) Perilipin 3 (PLIN3) protein content decreased with overfeeding (p=0.04, N=27). B) Baseline human primary myotubes treated with 200μM of palmitate showed that PLIN2 is expressed later over a time course of 2 hours. C) Changes in PLIN2 protein content correlated inversely with glucose disposal rate (GDR, r=-0.43, p=0.03, N=25). Graphs represent mean±S.E.M, *p<0.05.

Article Snippet: Antibodies against Total Akt (Cat no. 9272), p-S473 Akt (Cat no. 9271), Total IRS1 (Cat no. 2382S), p-S1101 IRS1 (Cat no. 2385), and mTOR (Cat no. 4517) were obtained from Cell Signaling Technology (Danvers, MA) The antibodies for PLIN2 (Cat no. NB110-40877) and PLIN3 (Cat no. NB110-40764) were obtained from Novus Biologicals (Littleton, CO).

Techniques: