Journal: Adipocyte

Article Title: Super-enhancer-associated miR-1260b coordinates adipogenesis and metabolic programming in human adipose stem cells

doi: 10.1080/21623945.2026.2638071

Figure Lengend Snippet: miR-1260b acts as a negative regulator of adipogenic differentiation. (A) qRT-PCR analysis validating the transfection efficiency, showing robust overexpression (OE) of miR‑1260b in hASCs compared with the normal control (NC). (B) Representative images of Oil Red O staining in hASCs transfected with NC, miR‑1260b mimic, or miR‑1260b inhibitor during adipogenesis. Scale bars represent 100 µm. (C, D) qRT-PCR (C) and Western Blot (D) analyses of key adipogenic markers ( PPARG , ADIPOQ , and FABP4 ) in the three groups. Data represent the mean ± SD. Significance was determined by unpaired two-tailed Student’s t test analysis. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The primary antibodies used were against PPARG (Cat: #2430S, Cell Signalling), ADIPOQ (Cat: #2789S, Cell Signalling), FABP4 (Cat: #50699S, Cell Signalling), and GAPDH (Cat: #10494, Proteintech).

Techniques: Quantitative RT-PCR, Transfection, Over Expression, Control, Staining, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation

doi: 10.1038/s41467-026-69196-4

Figure Lengend Snippet: A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of Ikaros binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and lenalidomide (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).

Article Snippet: Homogenates were subjected to SDS-PAGE, and blots were probed with primary antibodies against adiponectin (CST, #2789), beta-tubulin (CST, #2128), beta-actin (CST, #4970), vimentin (Sigma, #SAB4200716), glut4 (Fisher Scientifics, # MA183191 ), SerpinB2 (Invitrogen, #PA5-27857), Akt (CST, #4691), p-Akt ser 473 (CST, #4060), and GAPDH (CST, #2118).

Techniques: Imaging, Cell Culture, Staining, Expressing, Confocal Microscopy, Flow Cytometry, Western Blot, Biomarker Discovery, Binding Assay, RNA Sequencing, MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation

doi: 10.1038/s41467-026-69196-4

Figure Lengend Snippet: A Oxygen consumption rate (OCR) was measured by Seahorse XF 96 Extracellular Flux Analyzer (n = 20/group) in BMDM isolated from lean and obese mice. The correlation between maximal OCR and cytokine levels measured by ELISA in BMDM supernatants is shown. B GTT, ITT, and fasting insulin levels in HFD-fed LysM +/+ COX10 fl/fl and LysM cre/+ COX10 fl/fl mice (n = 7 for WT and 8 for KO, combined data of two independent experiments). C The Seahorse traces and maximal OCR (n = 24/group) in BMDM of obese SerpinB2 − / − or SerpinB2 +/+ mice are shown. D mRNA and protein levels of pro- and anti-inflammatory cytokines by qPCR and ELISA, respectively, were determined after SerpinB2 overexpression in THP-1 macrophages (n = 4/group, each dot represents cells cultured in one well.). E GTT and fasting insulin levels (n = 6–8/group, combined data of two independent experiments) (n = 8 for WT and 9 for KO) in obese SerpinB2 +/+ and SerpinB2 − / − mice. F – L HFD-fed LysM +/+ SerpinB2 fl/fl and LysM cre/+ SerpinB2 fl/fl mice were injected with PBS-vehicle, and LysM cre/+ SerpinB2 fl/fl mice were injected with IL-4. F GTT and ITT were performed, and the concentrations of fasting blood glucose, serum insulin, triglycerides (TG), free fatty acids (FFA), and free glycerol (FG) were evaluated (n = 10 for WT+vehicle, 18 for KO+vehicle, and 11 for KO + IL-4, combined data of at least two independent experiments). G Glucose infusion rate (GIR), hepatic endogenous glucose production (EGP), plasma insulin, and FFA levels are determined by hyperinsulinemic-euglycemic clamp studies (n = 7–9/group). Immunoblots ( H , I ) show pAkt, total Akt, Glut-4, adiponectin, and Ppar y expression in muscle and VAT (n = 5-6/group). The expression of the metabolic and inflammatory genes is measured by qPCR in whole VAT ( J ) (n = 8/group) and sorted VAT macrophages ( K ) (n = 6/group). L The expression of the anti-inflammatory genes was determined by RNA sequencing in CX 3 CR 1 + CCR2 + and CX 3 CR 1 − CCR2 − VAT macrophages isolated from lean mice (n = 3/group). Mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann–Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 6F, J).

Article Snippet: Homogenates were subjected to SDS-PAGE, and blots were probed with primary antibodies against adiponectin (CST, #2789), beta-tubulin (CST, #2128), beta-actin (CST, #4970), vimentin (Sigma, #SAB4200716), glut4 (Fisher Scientifics, # MA183191 ), SerpinB2 (Invitrogen, #PA5-27857), Akt (CST, #4691), p-Akt ser 473 (CST, #4060), and GAPDH (CST, #2118).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Over Expression, Cell Culture, Injection, Clinical Proteomics, Western Blot, Expressing, RNA Sequencing, MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation

doi: 10.1038/s41467-026-69196-4

Figure Lengend Snippet: A , B BODIPY images, adipocyte diameters and frequencies, and the expression of VAT expansion genes are shown in obese LysM cre/+ / Chr2 fl/fl mice exposed to either white (WLE) or blue (BLE) light (n = 8/group), and obese LysM +/+ SerpinB2 fl/fl and LysM cre/+ SerpinB2 fl/fl mice treated with vehicle and LysM cre/+ SerpinB2 fl/fl mice treated with IL-4 (n = 7/group). Scale bar=20 µm for BODIPY and bright field images, and 200 µm for VAT images. C Immunoblot images of the proteins in VAT of obese LysM cre/+ / Chr2 fl/fl , LysM +/+ SerpinB2 fl/fl , and LysM cre/+ SerpinB2 fl/fl mice (n = 5–6/group). D Confocal imaging to show the association between CCR2 − macrophages and adipocyte size in mouse VAT (n = 16/group). Scale bar =20 µm. E BODIPY and bright field images using ImageStreamX Mark II and Oil Red O pictures of undifferentiated or differentiated 3T3L1 cells cultured in the presence or absence of IL-4 or NAC (n = 7/group). Scale bar = 5 µm for the ImageStreamX images and 200 µm for the Oil Red O pictures. F The metabolic and inflammatory genes were measured by qPCR in 3T3 L1 differentiated adipocytes exposed to vehicle or NAC (n = 8/group). G – J Obese LysM cre/+ SerpinB2 fl/fl mice were supplemented with either water or NAC-containing water. G GTT and ITT were performed, and the concentrations of fasting blood glucose, serum insulin, triglycerides (TG), free fatty acids (FFA), and free glycerol (FG) were evaluated. (n = 8/group). H Immunoblots showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expression in muscle and VAT (n = 6/group). I The expression of the metabolic and inflammatory genes was analyzed in VAT by qPCR. (n = 12/group). J Quantification of CCR2 − macrophages in VAT. (n = 14/group). I , J Each dot represents one mouse. Mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 7B). The Mann–Whitney test (two-tailed) was used to determine the significance between two groups.

Article Snippet: Homogenates were subjected to SDS-PAGE, and blots were probed with primary antibodies against adiponectin (CST, #2789), beta-tubulin (CST, #2128), beta-actin (CST, #4970), vimentin (Sigma, #SAB4200716), glut4 (Fisher Scientifics, # MA183191 ), SerpinB2 (Invitrogen, #PA5-27857), Akt (CST, #4691), p-Akt ser 473 (CST, #4060), and GAPDH (CST, #2118).

Techniques: Expressing, Western Blot, Imaging, Cell Culture, MANN-WHITNEY, Two Tailed Test

Journal: PeerJ

Article Title: Capg enhances proliferation, adipogenesis, and inflammatory response in preadipocytes: insights from bioinformatics analysis and functional validation

doi: 10.7717/peerj.20730

Figure Lengend Snippet: (A) Representative images of Oil Red O staining in control (Ctrl) and CAPG-overexpressing ( Capg -OE) adipocytes on day eight of differentiation, visualizing lipid accumulation. (B) Quantitative analysis of the Oil Red O staining shown in (A) by elution and spectrophotometry ( n = 3 independent experiments). (C) High-magnification (200×) microscopic fields of the Oil Red O staining from (A); scale bar = 100 µm. (D) Quantitative analysis of the lipid droplet-positive area from the microscopic images in (C) ( n = 3 independent experiments). (E) Representative Western Blots analyzing the protein levels of CAPG and key adipogenic markers (PPARγ, adiponectin) in Ctrl and Capg-OE cells. (F–I) Densitometric quantification of the protein levels from (E) for (F) CAPG, (G) PPARγ, (H) PPARγ2, and (I) adiponectin ( n = 6 independent experiments). Data are presented as mean ± SD. Statistical significance was determined by Student’s t -test. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Primary antibodies used in this study included CAPG (1:2000, 10194-1-AP, Proteintech, Rosemont, IL, USA), β-tubulin (1:6000, 10094-1-AP, Proteintech, Rosemont, IL, USA), DDDDK-Tag (ABclonal, AE169PM, 47kD), PPARγ (1:1000, #2435T, CST), Adiponectin (1:1000, #2789, CST), CylinD1(1:2000, HY- P80098 , MCE).

Techniques: Staining, Control, Spectrophotometry, Western Blot