Fig 1: Protein levels of the selected candidates were examined in an independent validation cohort. (A, B) Representative Western blots and quantitative analysis showing the expression levels of PSMB9, CSTB, PRB4, DLAT, and F11 in plasma samples from 16 healthy controls, 16 non-RPP patients and 16 RPP patients. (C) Concentrations of CSTB and F11 in plasma were measured by ELISA from 16 healthy controls, 30 non-RPP patients and 30 RPP patients. Data are presented as mean ± SD. *p < 0.05 versus the healthy controls, #p < 0.05 versus the non-RPP patients.
Fig 2: Verification of candidate biomarkers in an in vitro RPP-like model. MH-S macrophages were treated with or without crystalline silica (CS) at 25 or 100 μg/cm² for 24 h. (A) Representative bright-field images showing morphological changes in MH-S cells following CS exposure. Scale bar, 100 μm. Cell viability (B), proliferation (C), and PI uptake (D) were assessed using MTT, CCK-8, and flow cytometry assays, respectively. (E) Levels of IL-1β, TNF-α, and IL-6 in culture supernatants were measured by ELISA. NIH-3T3 fibroblasts were then incubated with conditioned medium (CM) collected from MH-S cells treated as described above. (F) Representative images of wound healing assays at 0 and 24 h after CM treatment. Scale bar, 250 μm. (G) Quantitative analysis of wound closure rates. (H) Fibroblast proliferation was evaluated by CCK-8 assay following CM exposure. (I) Western blot analysis of COL1A1 and α-SMA expression in NIH-3T3 cells after CM treatment, with quantification shown in (J). (K) Representative Western blots and (L) quantification of PSMB9, CSTB, PRB4, DLAT, and F11 expression in NIH-3T3 cells. Data are presented as mean ± SD from at least three independent experiments. *p < 0.05 versus control group; #p < 0.05 versus 25 μg/cm² CS group.
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