Fig 1: Protein expression levels of MDA5, MAVS, IRF7, and ISG15 in the skeletal muscle of the anti‐MDA5 patients were significantly higher than those in the healthy controls (A, B) but lower than those in the antibody‐negative DM patients (C, D). Protein expression levels were quantified by densitometric analysis. Actin bands were used as loading control. Data are presented as mean ± SD. Statistical analysis was performed by Student’s t‐test. *P < 0.05, **P < 0.01.
Fig 2: Roles of IRF7/CH25HC/25HC in regulating macrophage polarization. (A) FC evaluated the role of diverse concentrations of 25HC in modulating macrophage polarization. N = 5. (B) qRT-PCR is used to evaluate the expression levels of diverse M1/M2 markers among diverse concentrations of 25HC groups. N = 4. (C) Effect of knockdown of IRF7 and CH25H expression levels in MB231 cells on M2 ratio using FC. N = 4. (D) Effect of knocking down the expression levels of IRF7 and CH25H on the end product 25HC using ELISA. N = 3. (E) Additional 25HC reverses the altered ability of MB231 to regulate macrophage polarization. N = 4. The data are evaluated with the 2-way and one-way ANOVA with the Tukey test or 2-tailed unpaired t-test. *, p<0.05; **, p<0.01; ***, p<0.005; ****, p<0.001.
Fig 3: Statuses of IRF7/CH25H/25HC pathway in diverse molecular types. (A) DEG identification based on the GSE42568 dataset. (B) The overlapping genes between DEGs and IRGs. (C) PPI network constructed by 268 overlapping genes. (D) The most significant module sorted out from the PPI network and potential macrophage-related biological process enriched by the 1 framed gene. (E) The different concentrations of 25HC among diverse cell lines. N = 3. (F) qRT-PCR analysis of IRF7 and CH25H among diverse cell lines. N = 6. (G) WB analysis of IRF7 among diverse cells. N = 3. (H) IHC analysis of the expression of IRF7 among diverse molecular types using BRCA samples. The total number of samples was 107 (HER2 positive n=15; Luminal A n=19; Luminal B n=17; TNBC n=24; non-cancer n=32). The data are evaluated with the 2-way and one-way ANOVA with the Tukey test or 2-tailed unpaired t-test. *p<0.05; **p<0.01; ***p<0.005; ****p<0.001.
Fig 4: Role of IRF7/CH25H/25HC on the proliferative capacity of breast cancer. (A) CCK-8 assay is used to measure the proliferative capacity of diverse cell lines with diverse concentrations of 25HC. N = 3. (B) FC assay for the effect of 25HC on the TNBC cell cycle. N = 4. (C) Inhibition of IRF7 expression status in TNBC cells inhibits tumorigenicity in vivo. N = 8. (D) Intervention with 25HC significantly reverses the slowing of tumor growth brought about by reduced IRF7. N = 4. (E) Inhibition of Irf7 expression status in 4T1 cells inhibits tumorigenicity in vivo. N = 4. (F) CD163+ percentage in TME between shRNA-Irf7-4T1 and shRNA-NC-4T1. N = 4. The data are evaluated with the 2-way and one-way ANOVA with the Tukey test or 2-tailed unpaired t-test. *p<0.05; **p<0.01; ***p<0.005.
Supplier Page from Abcam for Anti-IRF7 antibody