Fig 2: CircTMEM59 inhibited PDAC progression by regulating miR-147b/SOCS1 axis. (A) After transfection with si-SOCS1, the relative SOCS1 expression in PANC-1 and BXPC-3 cell lines were detected by Western blot analysis; **p < 0.01 vs. control, (B) Cell proliferation was detected by CCK8 assay in cells transfected with vector, circTMEM59, circTMEM59+ miR-147b or circTMEM59+ si-SOCS1; **p < 0.01 vs. vector group, #p < 0.05 vs. circTMEM59 group; (C) Cell invasion was detected by Transwell assay in cells transfected with vector, circTMEM59, circTMEM59+ miR-147b or circTMEM59+ si-SOCS1; **p < 0.01 vs. vector group, #p < 0.05 vs. circTMEM59 group; (D, E) The expression levels of SOCS1, PCNA, E-cadherin and vimentin were detected by Western blot analysis in cells transfected with vector, circTMEM59, circTMEM59+ miR-147b or circTMEM59+ si-SOCS1. PDAC, pancreatic ductal adenocarcinoma; **p < 0.01 vs. vector group, #p < 0.05 vs. circTMEM59 group. SOCS1, suppressor of cytokine signaling 1; PCNA, proliferation cell nuclear antigen.

Fig 3: SOCS1 was a target of miR-147b. (A) TargetScan predicted that SOCS1 was the potential target of miR-147b; (B) Luciferase assay confirmed that SOCS1 targeted miR-147b; **p < 0.01 vs. miR NC; (C) After transfection with miR-147b, the relative SOCS1 expression in PANC-1 and BXPC-3 cell lines were detected by Western blot analysis; **p < 0.01 vs. miR NC; (D) After transfection with circTMEM59 or circTMEM59 + miR-147b, the relative SOCS1 expression in PANC-1 and BXPC-3 cell lines were detected by Western blot analysis. **p < 0.01 vs. vector, #p < 0.05 vs. circTMEM59; (E) The SOCS1 expression in PDAC tumor tissues and adjacent normal tissues was measured by qRT-PCR; ***p < 0.001 vs. NC; (F) Correlation analysis of the expression of SOCS1 and miR-147b in 66 PDAC tissues; (G) Correlation analysis of the expression of SOCS1 and circTMEM59 in 66 PDAC tissues. SOCS1, suppressor of cytokine signaling 1; PDAC, pancreatic ductal adenocarcinoma.

Fig 4: Differential effects of aged and infected PMNs on intracellular M1 markers and cytokine secretion. (A) Lysates of M0, M1 and M2 macrophages and M1 macrophages that ingested aged or infected PMNs (upper left and lower left blots) or M1 macrophages directly infected with F. tularensis (upper right) were probed to detect TGM2, IDO and SOCS1 with β-tubulin as the loading control. Data from three independent experiments are shown (n1-n3). (B-E) Secretion of TNFα (B), MIP-1α (C), IL-18 (D) and IL-1β (E) were measured by ELISA. Data are the mean ± SD from three independent experiments and were analyzed by two-way ANOVA with Tukey’s multiple comparisons post-test. *p < 0.05, **p < 0.01, ****p < 0.0001.

Fig 5: Proliferation, migration and invasion capabilities of A375 cells after knockdown of IFNG, PTPN6, SLC38A1 and SOCS1. (A, B) Protein and mRNA expression levels of IFNG, PTPN6, SLC38A1, and SOCS1 after siRNA transfection. (C) Proliferative capacity of A375 cells after knockdown of IFNG, PTPN6, SLC38A1, and SOCS1 via CCK8 assay. (D) Transwell invasion assay for detection of invasive capacity of A375 cells after knockdown of IFNG, PTPN6, SLC38A1 and SOCS1. (E) Wound healing assay for detection of migratory capacity of A375 cells after knockdown of IFNG, PTPN6, SLC38A1, and SOCS1, ns:not significant, *p< 0.05, **p< 0.01, ***p< 0.001.