Journal: International Journal of Biological Sciences

Article Title: Atrial Natriuretic Peptide Attenuates Colitis via Inhibition of the cGAS-STING Pathway in Colonic Epithelial Cells

doi: 10.7150/ijbs.67356

Figure Lengend Snippet: Expression of atrial natriuretic peptide (ANP) and its receptor in colon and serum samples. (A) Relative mRNA levels of the ANP precursor (NPPA) and ANP receptors (NPR-A and NPR-C) in different organs of wild-type mice as determined by quantitative real-time polymerase chain reaction (qRT-PCR; n = 5 for each group). The Y axis represents the expression level difference between the cycle threshold (CT) value of the target gene and the β-actin, and the smaller the Y axis value, the higher the gene expression. (B) Mice were treated with 3% dextran sulfate sodium (DSS) in drinking water for 7 days. Relative expression of NPPA, NPR-A , and NPR-C in the colon of control and DSS-treated groups was detected by qRT-PCR. Results were normalized against the β-actin gene. (C) Level of ANP was determined in the serum of a DSS-induced colitis and control mice using an enzyme-linked immunosorbent assay (ELISA). (D) Relative expression of NPPA , NPR-A , and NPR-C in the colon of people with UC in activity (n = 15), UC in remission (n = 9) and control individuals (n = 45) was determined using RT-qPCR. (E) Level of ANP was detected in serum from UC patients with active disease (n = 12) and remission (n = 11) and control individuals (n = 27) using ELISA. (F) Correlation between murine serum ANP and percent weight loss. (G-I) Correlation between serum ANP and C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR) and neutrophilicgranulocyte (NE) level. (J) NPR-A expression was detected by immunohistochemical analysis in murine colonic tissue. (K) The level of adrenaline was determined using ELISA with the treatment of DSS and ANP or not. (L) The level of adrenaline was detected in serum from people with UC or controls using ELISA. (M) Mice with a DSS-induced experimental model of colitis (n = 6 in each group) were intraperitoneally injected with prazosin or PBS. Bodyweight curves were recorded. Statistical analysis was performed using T-tests: NS (not significant), P > 0.05; *P < 0.05; **P < 0.01; and ****P < 0.0001 are shown on the figure.

Article Snippet: Meanwhile, prazosin (0.2 mg in 400 μL phosphate buffered saline [PBS] per mouse; Xinyi Pharmacy, China), human ANP recombinant protein (2 μg in 400 μL PBS per mouse; Tocris Bioscience, England), STING pathway agonist DMXAA (0.1 mg in 400 μL PBS per mouse; Topscience, China), or an equal volume of PBS as a control was injected intraperitoneally into the mice at the same time daily.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Gene Expression, Control, Enzyme-linked Immunosorbent Assay, Activity Assay, Sedimentation, Immunohistochemical staining, Injection

Journal: Cell Death & Disease

Article Title: FAM3A drives uncoupling of muscle lipid accumulation and insulin resistance depending on insulin receptor

doi: 10.1038/s41419-025-08298-1

Figure Lengend Snippet: a Western blot images showing the presence of FAM3A in different cellular compartments in soleus muscles and heart tissues. b Levels of the FAM3A protein along with positive controls (TSG101 and CD9) and the negative control (calnexin) in plasma-derived exosomes from patients chosen at random. c Western blot images and quantification of the levels of p-AKT (thr308) and p-insR-β (tyr1150/1151) in cells treated with different concentrations (ng/ml) of rcFAM3A for 8 h (n = 4 biologically independent samples/group; quantitative comparisons between samples were run on the same gel). d Western blot images and quantification of the expression levels of the indicated proteins in soleus muscles from the mice treated with bms536924 and fed an HFD for 10 weeks (n = 6 biologically independent animals/group; quantitative comparisons between samples were run on the same gel). e Western blot images and quantification of the expression levels of the indicated proteins in cells pre-treated with bms536924 or transfected with the insR siRNA and then stimulated with rcFAM3A (200 ng/ml; n = 4 biologically independent samples/group; quantitative comparisons between samples were run on the same gel). The lipid droplets detected by oil red O staining ( f ) and TG content ( g ) were measured in soleus muscles from the mice treated with bms536924 and fed an HFD for five weeks and the results were plotted (n = 6 biologically independent animals/group). Scale bar: 200 μm, insets: 100 μm in ( f ). C2C12 cells were treated with rcFAM3A (200 ng/ml) for 12 hours. Cellular De novo FA synthesis ( h ) and TG content ( i ) were measured and graphed (n = 3 biologically independent samples/group). j , k Body adiposity and body weight gain were measured in mice treated with bms536924 and fed an HFD for 10 weeks and the results were plotted (n = 7 biologically independent animals/group). The data are presented as the means ± SEMs. Statistical significance was determined with a two-tailed independent t test, and P values are indicated ( ns P ≥ 0.05). Source data are provided as a file.

Article Snippet: For the experiments, the C2C12, HL-1, and 3T3-L1 cell lines were treated with 200 ng/ml rcFAM3A or chemical PPARα inhibitor GW6471 (8 μM diluted with DMSO; Topscience, #T8486) [ ] or chemical insulin receptor (insR) inhibitor bms536924 (4 μΜ diluted with DMSO; Topscience, #T6419) for 12 h. For insR silencing, C2C12 cells were incubated with 0.5 μg/ml siRNA (Santa Cruz, #sc-35673).

Techniques: Western Blot, Muscles, Negative Control, Clinical Proteomics, Derivative Assay, Expressing, Transfection, Staining, Two Tailed Test