Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Grass carp can adapt to short-term HFD by expanding healthy adipose tissue . A , body weight from grass carp–fed Control (normal diet) and HFD (high-fat diet) for 3 days (D3), 2 weeks (W2), 4 weeks (W4), and 8 weeks (W8), n = 3. B , hepatosomatic from grass carp–fed Control and HFD for D3, W2, W4, and W8. Hepatosomatic index (HSI, %) = 100 × (liver weight, g)/(individual fish weight, g), n = 3. C , intraperitoneal fat index from grass carp–fed Control and HFD for D3, W2, W4, and W8. Intraperitoneal fat index (IPFI, %) = 100 × (intraperitoneal fat weight, g)/(final weight, g), n = 3. D , H&E stains of the adipose tissue from grass carp–fed Control and HFD for D3, W2, W4, and W8. Scale bar represents 50 μm. E , using ImageJ software to measure relative adipocyte size combined with H&E staining results of the adipose tissue, n = 3. F , representative images of immunofluorescence staining for EdU in the adipocyte nuclei of adipose tissue from grass carp after 1 week of EdU treatment and 2 weeks on the indicated diet. Tissue is also stained for caveolin-1 ( green ) to visualize adipocyte plasma membranes, as well as DAPI ( blue ) to visualize nuclei. Scale bar represents 50 μm. G , quantitative analysis of EdU positive nuclei in grass carp adipocytes after 3 days, 2 weeks, 4 weeks, and 8 weeks of conditional diet. The yellow arrows indicate edu-positive adipocyte nuclei, which are located within the cytoplasmic membrane of adipocytes, n = 3. H , growth rate of EDU-positive cell nuclei in adipocyte at different time periods, n = 3. I , immunohistochemical staining with an F4/80 antibody of adipose tissue, crown-like structures (CLS); scale bar represents 50 μm. J , quantification of CLS, n = 3. K , the mRNA expression of IL-1β, IL-8, TNF-α in adipose tissue, n = 3. Results are presented as means ± SD. Data analysis was conducted by Student’s two-tailed t test. Results with p < 0.05 represents a statistically significant difference: ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: For immunofluorescence staining, adipose tissue paraffin sections were blocked with bovine serum albumin and incubated overnight with F4/80 antibody (1:200, bs-11182R, Bioss) at 4 °C to specifically bind to macrophages.

Techniques: Control, Software, Staining, Immunofluorescence, Clinical Proteomics, Membrane, Immunohistochemical staining, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Inhibition of adipocyte hyperplasia and induction of pathological expansion of adipose tissue under short-term HFD conditions . A , body weight of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. B , intraperitoneal fat index of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. C , anatomical diagram of abdominal adipose tissue of grass carp and its H&E staining; scale bar represents 50 μm. D , triglyceride content in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. E , relative adipocyte size in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. F , relative adipocyte number in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. G , quantitative analysis of EDU-positive cell nuclei in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. H , immunohistochemical staining with an F4/80 antibody of adipose tissue, crown-like structures (CLS); scale bar represents 50 μm. H ′, quantification of CLS, n = 3. I , the mRNA expression of IL-1β, IL-8 in adipose tissue, n = 3. J , Western blot analysis of PPARγ protein levels in isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, HFD + PX-478, and HFD + GW9662. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. J ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. K , Western blot analysis of PPARγ protein levels in isolated SVF obtained directly from grass carp adipose tissue after 3 days of Control and HFD. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. K ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. Data are represented as the mean ± SD. For comparisons between two groups, a two-tailed Student’s t test was performed, ∗ p < 0.05. Statistical testing was performed with one-way ANOVA for comparisons among three or more groups. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: For immunofluorescence staining, adipose tissue paraffin sections were blocked with bovine serum albumin and incubated overnight with F4/80 antibody (1:200, bs-11182R, Bioss) at 4 °C to specifically bind to macrophages.

Techniques: Inhibition, Staining, Immunohistochemical staining, Expressing, Western Blot, Isolation, Control, Two Tailed Test

Journal: bioRxiv

Article Title: Macrophage Antigen Presentation Is Unleashed by Pan-RAS Inhibition to Promote Antitumor Immunity

doi: 10.64898/2026.01.30.702886

Figure Lengend Snippet: A) Experimental scheme for B16-OVA transplantation in nude mice followed by RMC-6236 oral gavage (10 mg/kg, daily) ; B) C omparison of tumor size between vehicle and RMC-6236–treated nude mice ; C) Tumor growth curves ; D,E) Flow-cytometry analysis of TAM CD206 (D) and CD86 (E) with vehicle or RMC-6236 ; F) Immunofluorescence showing macrophages (F4/80, green) and MHC-I (red) with DAPI (blue) ; G–I) Flow-cytometry quantification in TAMs of MHC-I (H-2K b ) (G) , OVA-specific presentation (SIINFEKL) (H) , and MHC-II (I-A/I-E) (I); J) Schematic of anti-CSF1R– mediated monocyte/macrophage depletion during RMC-6236 treatment; K) Tumor growth with RMC-6236 plus IgG or anti-CSF1R ; L, M) Intratumoral CD8 + T-cell frequency after monocyte depletion ; N,O) Granzyme B (N) and perforin (O) in tumor CD8 + T-cells. Data are shown as mean ± SEM. *p < 0.05, **p< 0.01, ***p < 0.001 and ****p < 0.0001 using unpaired two-tailed t-tests and Two-way ANOVA followed by Sidak’s multiple comparisons test (n≥6).

Article Snippet: Macrophages were purified using anti-mouse F4/80 MicroBeads (Miltenyi Biotec) according to the manufacturer’s protocol.

Techniques: Transplantation Assay, Flow Cytometry, Immunofluorescence, Two Tailed Test