Fig 1: ASM exhibits high expression in DM patients and associates with disease activity. (A) serum ASM levels detected using ELISA (B) mRNA expression level of ASM in muscle tissues of patients (C) protein levels of ASM in patients’ muscle tissues, scale bar 20 μm (D) serum ASM activity (E) Correlation analysis between serum ASM levels and overall disease activity (F) Correlation analysis between serum ASM levels and cutaneous severity score. *P < 0.05, **P < 0.01, ***P < 0.001, n = 30 in healthy controls, n = 58 in patients with DM
Fig 2: The proportion of CD4 T cell subsets was imbalanced in EAM mice model. Flow cytometry was applied to test the CD4 T cell subsets Treg and Th17 in spleen. (A) Treg in the wild type EAM model mice (B) Th17 in the wild type EAM model mice (C) Treg in the SMPD1−/− EAM model mice (D) Th17 in the SMPD1−/− EAM model mice (E) Th17 in the wild type EAM model mice treated with amitriptyline (F) Treg in the wild type EAM model mice treated with amitriptyline (G) Treg ratio of CD25 + Foxp3 + versus CD25-Foxp3 + in SMPD1−/− EAM model mice (H) Treg ratio of CD25 + Foxp3 + versus CD25-Foxp3 + in EAM model mice treated with amitriptyline. *P < 0.05, ** P < 0.01, *** P < 0.001, n ranges from 5 to 7 in each analysis group
Fig 3: ASM plays a crucial role in the pathology of EAM mice. Establishment of EAM mouse model in ASM knockout (SMPD1−/−) mice. (A) muscle strength (B) serum CK level (C) spleen (D) spleen weight (E) H&E staining of muscle section (F) quantitative analysis of H&E staining. *P < 0.05, ** P < 0.01, *** P < 0.001, n = 6 in SMPD1+/+ mice group, n = 5 SMPD1−/− mice group
Fig 4: Amitriptyline is therapeutic against EAM mice model. To establish the EAM mouse model, wild type mice were subject to induction and subsequently treated with amitriptyline (AMI) at dosage levels of 1 mg/kg or 10 mg/kg. (A) muscle strength (B) serum CK level (C) spleen (D) spleen weight (E) H&E staining of muscle section (F) quantitative analysis of H&E staining. (G) mRNA expression level of ASM in muscle tissues (H) serum ASM activity. *P < 0.05, ** P < 0.01, *** P < 0.001, n ranges from 3 to 7 in each analysis group
Supplier Page from Abcam for Human Acid sphingomyelinase ELISA Kit (SMPD1)