Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: METTL1-deficient MSCs inhibit lipid synthesis in hepatocytes. (A) Representative images of Nile Red staining in hepatocytes co-cultured with MSC shGFP and MSC shMETTL1 following treatment with FFA (Scale bar = 20 μm). (B) Measurement of TG content in hepatocytes in the indicated groups. (C, D) Western blot analysis of lipid metabolism-related gene expression (FASN, SREBP1, SCD1) in the indicated groups. (E, F) qPCR analysis of lipid synthesis gene expression ( Fasn, Scd1, Srebp1, Fads1 and Acaca ) in AML12 and HepG2 cells co-cultured with MSC shGFP and MSC shMETTL1 . For all statistical graphs, data are presented as mean ± S.E.M, with statistical significance is indicated in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: Staining, Cell Culture, Western Blot, Gene Expression

Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: Transplantation of METTL1-deficient MSCs alleviates metabolic disorders associated with MASLD. (A) Schematic diagram of the animal experiment. (B) Evaluation of liver weight and the liver-to-body weight ratio in the indicated mice. (C) Assessment of fasting blood glucose levels in the indicated mice. (D) Analysis of GTT and ITT for the indicated groups. (E) Measurement of serum ALT and AST levels following 7 weeks of cell transplantation. (F) Representative images of HE and Oil Red O staining for analysis of mouse liver tissue (Scale bar = 100 μm). (G) Determination of TG and TC levels in the liver tissue of the specified mice. (H) qPCR analysis of lipid synthesis-related genes, including Fasn, Scd1, Srebp1, Fads1 , and Acaca in the specified groups. (I) Western blot analysis of lipid metabolism-related proteins in the specified groups. For all statistical graphs, individual data points represent individual mice, and data are presented as mean ± S.E.M. Statistical significance is indicated as shown in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: Transplantation Assay, Staining, Western Blot

Journal: Stem Cells Translational Medicine

Article Title: METTL1-deficient mesenchymal stem cells protect against metabolic-associated fatty liver disease by increasing NAMPT secretion

doi: 10.1093/stcltm/szag016

Figure Lengend Snippet: NAMPT/SIRT1/SREBP1 mediates the protective effects of METTL1-deficient MSCs in MASLD. (A-C) Representative images of IF staining for NAMPT, SIRT1 and SREBP1 in HepG2 cells co-cultured with MSC shGFP and MSC shMETTL1 cells after treatment with FFA (Scale bar = 20 μm). (D, E) Western blot analysis of NAMPT, SIRT1 and SREBP1 expression in HepG2 cells co-cultured with MSC shGFP and MSC shMETTL1 cells following FFA treatment. (F) The NAD+ content was measured in the indicated cells. (G-I) Representative images of IF staining for NAMPT, SIRT1 and SREBP1 in liver tissues from mice transplanted with MSC shGFP and MSC shMETTL1 cells after 16 weeks of HFD feeding (Scale bar = 100 μm). (J, K) Western blot analysis of NAMPT, SIRT1, and SREBP1 expression in liver tissues from mice transplanted with MSC shGFP and MSC shMETTL1 cells after 16 weeks of HFD feeding. (L) The NAD+ content was measured in the indicated mouse liver tissues. For all statistical graphs, data are presented as mean ± S.E.M., with statistical significance indicated in the figure.

Article Snippet: Additionally, a primary antibody to METTL1 (14994-1-AP), SIRT1 (13161-1-AP), NAMPT (11776-1-AP) and SREBP1 (14088-1-AP) were purchased from Proteintech.

Techniques: Staining, Cell Culture, Western Blot, Expressing

Journal: Journal of Traditional and Complementary Medicine

Article Title: Integration proteomics analysis to identify AMPK as key target pathways of TCM formula for high fat diet induced obesity in mice

doi: 10.1016/j.jtcme.2025.02.010

Figure Lengend Snippet: LAH regulates lipogenesis by AMPK signaling pathway. (A and B) The protein expressions of p-AMPK, AMPK, p-ACC, ACC, CPT1A, FAS, and SREBP1 were analyzed in the protein lysates from HepG2 and AML12 cells using Western blotting. Quantification of p-AMPK/AMPK, p-ACC/ACC, CPT1A/GAPDH, FAS/GAPDH and SREBP1/GAPDH ratios was performed using ImageJ. Data are represented as the mean ± SD (n = 3/group). ∗ p < 0.05 compared to the blank group. + p < 0.05 compared to OA group. (C) Immunofluorescence staining of SREBP1 and FAS expression in OA-induced AML12 cells. (D) Quantitative analysis of the expression of SREBP1 and FAS in immunofluorescence staining using ImageJ. Data are represented as the mean ± SD (n = 2/group). ∗ p < 0.05 compared to the blank group. + p < 0.05 compared to OA group.

Article Snippet: The protein targets related to fat metabolism were CPT1, SREBP1, p-AMPK, AMPK, ACC, and p-ACC (Cell Signaling Technology, Denvers, MA, USA).

Techniques: Western Blot, Immunofluorescence, Staining, Expressing

Journal: Animal Nutrition

Article Title: Influences of dietary guanidinoacetic acid supplementation on performance and proteins involved in milk fat and protein synthesis in dairy cows

doi: 10.1016/j.aninu.2025.07.008

Figure Lengend Snippet: Changes in the expression levels of proteins concerned with fatty acid synthesis. (A) Western blot analysis of AMPK, p-AMPK, PPARγ, SREBP1, ACACA, p-ACACA, FASN, and SCD1 in bovine mammary gland tissues treated with 0 g/kg (control) and 0.36 g/kg DM of GAA derived from RPGAA (MRPGAA); β-actin was used as the loading control. (B) Mean ± SEM of immunopositive bands of PPARγ, SREBP1, p-ACACA/ACACA, FASN, SCD1, and p-AMPK/AMPK. AMPK = adenosine monophosphate activated protein kinase; PPARγ = peroxisome proliferator-activated receptor gamma; SREBP1 = sterol regulatory element-binding protein 1; ACACA = acetyl-coenzyme A carboxylase-α; FASN = fatty acid synthase; SCD1 = stearoyl-CoA desaturase 1; GAA = guanidinoacetic acid; RPGAA = rumen-protected guanidinoacetic acid; p = phosphorylated. ∗ P < 0.05 and ∗∗ P < 0.01 versus the control group.

Article Snippet: Rabbit anti-sterol regulatory element-binding protein 1 (SREBP1; #NB100-2215) was purchased from Novus Biologicals (Centennial, CO, USA).

Techniques: Expressing, Western Blot, Control, Derivative Assay, Binding Assay