Fig 2: DHA inhibited the JAK2/STAT3 pathway in pneumonia tissue. (A) Tgfb1, (B) Jak2, and (C) Stat3 mRNA levels in the lungs were determined using qRT-PCR (n = 6), data are expressed as the fold change in mRNA expression normalized to Gapdh expression, with respect to the control group. Representative images of (D) TGF-ß1 and (E) JAK2 protein expression in lung tissue detected using IHC. Mean IOD of (F) TGFß1 and (G) JAK2 protein by IHC staining. Data are expressed as the means ± standard error. *P < 0.05, **P < 0.001. Abbreviations: BLM: bleomycin; DHA-1: dihydroartemisinin 25 mg/kg/d; DHA-2: dihydroartemisinin 50 mg/kg/d; DHA-3: dihydroartemisinin 100 mg/kg/d; IHC: immunohistochemistry; IOD: integrated optical density; NS: not statistically significant.

Fig 3: JAK2 or HIF-1 inhibition mitigated CALM2-mediated macrophage polarization and angiogenesis. The JAK2 inhibitor LY2784544 (1 μM) and HIF-1 inhibitor SYP-5 (10 μM) were deployed to treat AGS and MKN45 cells with CALM2 overexpression. The conditioned medium was collected and cultivated along with macrophages and HUVECs for 24 hours. (A) Western Blot was done to figure out the profile of the JAK2/STAT3/HIF-1/VEGFA axis in macrophages. (B, C) Western Blot also ascertained the protein profiles of CD206, CD163, CD11b, CD80, CD86, and iNOS in macrophages. (D) qRT-PCR disclosed the mRNA expressions of M2-type macrophage-concerned factors CXCL12, IL-4, IL-13, IL-10, and VEGFA. (E) Western Blot verified the profile of the JAK2/STAT3/HIF-1/VEGFA pathway in HUVECs. Data were presented as mean ± SD (n=3). NSP > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001 (vs. the CM+CALM2 group).

Fig 4: FNDC5 deficiency enhanced HFD-induced oxidative stress and phosphorylated JAK2 and STAT3 level in heart. a Superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in heart. b Expression of NAPDH oxidases (NOX2 and NOX4) protein in heart. c Phosphorylated JAK2 and STAT3 level in heart. Values are mean ± SEM. *P < 0.05 vs. WT. †P < 0.05 vs. Ctrl. n = 6

Fig 5: Expression of NF-?B, AICDA, Akt, JAK2/STAT3, and IL6 by lung adenocarcinoma cell lines. (A) Basal expression of NF-?B components, AICDA, p-Akt, Akt, p-Jak2, Jak2, p-Stat3, and Stat3, and relative expression of IL-6 mRNA, by the PC9 (adenocarcinoma with an EGFR exon 19 deletion) and PC9/GR (EGFR-TKI-resistant adenocarcinoma with the T790M mutation) cell lines. Changes in expression in (B) NF-?B components and (C) AICDA, p-Akt, Akt, p-Jak2, Jak2, p-Stat3, and Stat3, and in relative expression of IL-6, after treatment with gefitinib (EGFR-TKI). Uncropped Western blot films are listed in Supplementary Figure S2A–C. ß-actin was used as a loading control. Two-tailed Student’s t-test was used to determine statistical significance. A p-value < 0.05 was considered statistically significant (* p < 0.005).