Journal: Cancer & Metabolism

Article Title: The FGL1-LAG-3 axis attenuates melanoma-induced cachexia in mice

doi: 10.1186/s40170-025-00418-x

Figure Lengend Snippet: Fgl1 upregulation in WAT and liver in mice models of cancer cachexia. ( A ) Interventional design for B16F10 cachexia mouse model studies. ( B ) Tumor weight of B16F10 mice on day 18 after cell injection ( n = 3 per group). ( C ) Tumor-free body weight of PBS and B16F10 injected mice on day 18 ( n = 3 per group). ( D , E ) RNA-seq design and analysis ( n = 3 per group). ( F ) KEGG pathway analysis of RNA-seq data ( n = 3 per group). ( G ) mRNA expression of Fgl1 in eWAT. The expression of each gene was normalized to actin. ( n = 3 per group). ( H ) Protein expression of Fgl1 in eWAT. ( n = 3 per group). ( I ) Gene expression of FGL1 in the spleen, muscle, kidney, lung, liver, stomach, and intestine from the mice ( n = 3 per group). All values are presented as mean ± standard error of the mean (SEM). P values were determined by two-tailed Student’s t-test, with * p < 0.05, ** p < 0.01, and *** p < 0.001 indicating different levels of statistical significance compared to control. ns, not significant

Article Snippet: The B16F10 cell line was maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sangon Biotech, E600003) supplemented with 10% fetal bovine serum (FBS, Sangon Biotech, E600052) and 1% penicillin-streptomycin (P/S, Sangon Biotech, E607011).

Techniques: Injection, RNA Sequencing, Expressing, Gene Expression, Two Tailed Test, Control

Journal: Cancer & Metabolism

Article Title: The FGL1-LAG-3 axis attenuates melanoma-induced cachexia in mice

doi: 10.1186/s40170-025-00418-x

Figure Lengend Snippet: Early treatment with FGL1 was effective in promoting tumor growth, while intermediate and late-stage treatment exhibited little effect. ( A ) Experimental design of FGL1 treatment time point. ( B ) FGL1-fc injection on day 7 promotes B16F10 growth ( n = 5 per group). ( C ) FGL1-Fc injection on day 14 does not impact B16F10 growth ( n = 5 per group). All values are presented as mean ± SEM. P values were determined by one-way analysis of variance (ANOVA), with * p < 0.05, ** p < 0.01, and *** p < 0.001 indicating different levels of statistical significance compared to control. ns, not significant

Article Snippet: The B16F10 cell line was maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sangon Biotech, E600003) supplemented with 10% fetal bovine serum (FBS, Sangon Biotech, E600052) and 1% penicillin-streptomycin (P/S, Sangon Biotech, E607011).

Techniques: Injection, Control

Journal: Cancer & Metabolism

Article Title: The FGL1-LAG-3 axis attenuates melanoma-induced cachexia in mice

doi: 10.1186/s40170-025-00418-x

Figure Lengend Snippet: FGL1-fc treatment counteracts cancer cachexia by alleviating body weight loss and WAT and muscle wasting. ( A ) Study design for the FGL1-fc treatment. ( B ) Tumor-free body weight% change of B16F10 tumor-bearing mice treated with vehicle and FGL1-fc ( n = 5 per group). ( C ) Measurement of the food intake (the food intake of mice is calculated as the average of a cage of 5 mice). ( D ) Fat percentage of mice ( n = 5 per group). ( E ) Lean mass percentage of mice ( n = 5 per group). ( F – I ) Representative anatomical image, weight, hematoxylin and eosin ( H & E ) staining and adipocyte size of eWAT ( n = 5 per group). ( J – M ) Representative anatomical image, weight, H&E staining and cross-section area of gastrocnemius muscles ( n = 5 per group). All values are presented as mean ± SEM. P values were determined by one-way ANOVA, with * p < 0.05, ** p < 0.01, and *** p < 0.001 indicating different levels of statistical significance compared with control. ns, not significant

Article Snippet: The B16F10 cell line was maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sangon Biotech, E600003) supplemented with 10% fetal bovine serum (FBS, Sangon Biotech, E600052) and 1% penicillin-streptomycin (P/S, Sangon Biotech, E607011).

Techniques: Staining, Muscles, Control

Journal: Cancer & Metabolism

Article Title: The FGL1-LAG-3 axis attenuates melanoma-induced cachexia in mice

doi: 10.1186/s40170-025-00418-x

Figure Lengend Snippet: FGL1-fc reverses lipid and muscle metabolism alterations in B16F10 cachectic mice. ( A ) Three pathways of fatty acid metabolism. ( B ) FGL1-fc counteracts a decline in the expression of fatty acid synthesis genes FAS and SCD1 ( n = 5 per group). ( C ) Both B16F10 tumor and FGL1-fc do not impact the gene expression of lipolysis ( n = 5 per group). ( D ) FGL1-fc does not counteract the decrease in the expression of fatty acid storage gene DGAT2 ( n = 5 per group). ( E ) Levels of HSL or p-HSL in eWAT ( n = 4 or 5 per group). ( F , G ) Circulating esterified Free Fatty Acids (NEFA) and glycerol level ( n = 5 per group). ( H – J ) Skeletal muscle expression of atrophy-associated genes, Mustn1 and IGF1, is protected by FGL1-fc ( n = 5 per group). All values are presented as mean ± SEM. P values were determined by one-way ANOVA, with * p < 0.05, ** p < 0.01, and *** p < 0.001 indicating different levels of statistical significance compared with control. ns, not significant

Article Snippet: The B16F10 cell line was maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sangon Biotech, E600003) supplemented with 10% fetal bovine serum (FBS, Sangon Biotech, E600052) and 1% penicillin-streptomycin (P/S, Sangon Biotech, E607011).

Techniques: Expressing, Gene Expression, Control

Journal: Cancer & Metabolism

Article Title: The FGL1-LAG-3 axis attenuates melanoma-induced cachexia in mice

doi: 10.1186/s40170-025-00418-x

Figure Lengend Snippet: Anti-cachectic effect of FGL1 via LAG-3. ( A ) Experimental design of LAG-3 antibody studies. ( B ) Intermediate and late-stage treatment with LAG3 block antibody or FGL1-fc had no effect on B16F10 tumor growth ( n = 5 per group). ( C – E ) Representative body weight, eWAT weight, and gastrocnemius muscles weight following FGL-fc or FGL1-fc + anti-LAG-3 antibody treatment ( n = 5 per group). All values are presented as mean ± SEM. P values were determined by one-way ANOVA, with * p < 0.05, ** p < 0.01, and *** p < 0.001 indicating different levels of statistical significance compared to control. ns, not significant

Article Snippet: The B16F10 cell line was maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sangon Biotech, E600003) supplemented with 10% fetal bovine serum (FBS, Sangon Biotech, E600052) and 1% penicillin-streptomycin (P/S, Sangon Biotech, E607011).

Techniques: Blocking Assay, Muscles, Control

Journal: Aging Cell

Article Title: Liver osteopontin is required to prevent the progression of age‐related nonalcoholic fatty liver disease

doi: 10.1111/acel.13183

Figure Lengend Snippet: Liver OPN is p53 regulated. (a) Protein levels of p53 were evaluated by immunohistochemistry in liver sections of 3‐, 10‐, and 20‐month‐old (m) wild‐type (WT) mice ( n = 3–5). (b) OPN protein levels from liver homogenates were measured in WT and p53‐KO male and female mice fed a chow diet (CD) and a high‐fat diet (HFD) by immunoblotting using glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) as loading control ( n = 4–6). (c) Liver p53 and OPN protein levels were evaluated in WT mice fed a HFD injected with p53‐dominant positive adenovirus (adp53) and GFP (adGFP), using transferrin or GAPDH as a loading control ( n = 4–6). (d) Protein levels of p53 and OPN in siCtrl and sip53 HepG2 cells were measured by immunoblotting using GAPDH as loading control. Extracellular OPN was measured using an ELISA ( n = 4–5). (e) OPN protein levels from HepG2 cells silenced for p53 and Hep3B cells treated with palbociclib (Palbo) were measured by immunoblotting using GAPDH as loading control. OPN media levels from Hep3B cells treated either with vehicle (Veh) or with palbociclib (Palbo) were measured by ELISA ( n = 4–8). Values are means ± SEM . Significant differences are denoted by * p < 0.05, ** p < 0.01, and *** p < 0.001 (Student's t test)

Article Snippet: Data obtained from the Cell Line Encyclopedia (© 2019 The Broad Institute of MIT & Harvard) showed that the cell line with the least p53 expression, the Hep3B, also had the lowest OPN expression.

Techniques: Immunohistochemistry, Western Blot, Control, Injection, Enzyme-linked Immunosorbent Assay