Journal: Cellular and Molecular Immunology

Article Title: Synergistic effects of LCN2 and TWEAK on the progression of psoriasis

doi: 10.1038/s41423-025-01292-9

Figure Lengend Snippet: TWEAK, Fn14, and LCN2 are overexpressed in skin lesions of psoriasis patients and IMQ induced psoriatic mice model. A Single cell data from psoriasis was used for correlation analysis between TWEAK, Fn14 and LCN2 . B Immunostaining of LCN2, TWEAK and Fn14 in the skin of healthy control (HC) and patient with psoriasis (Brown). Normal control skin samples = 4, patient skin samples = 4. Nuclei were stained with hematoxylin (Bar = 200 μm). C Immunostaining of LCN2, TWEAK and Fn14 were performed on wild-type mouse skin paraffin sections. Stained epidermal region areas were quantitated by ImageJ software. The relationship between the IOD of TWEAK (or Fn14) and LCN2 staining was assessed by Spearman’s rank correlation. Number of control mice = 3, Number of IMQ induced WT mice = 4. Representative images are shown (Bar = 200 μm). D Volcano Plot showed the RNA-seq analysis of wild-type control mice and wild-type imiquimod model mice. E RNA-seq analysis of wild-type mice and Lcn2 –/– mice treated with imiquimod. F Immunostaining of TWEAK, Fn14 and Ly6G were performed on Lcn2 knockout mice skin paraffin sections. The relationship between the IOD of TWEAK (or Fn14) and number of Ly6G positive cell staining was assessed by Spearman’s rank correlation. Number of control mice = 3, Number of IMQ induced WT mice = 4, Number of IMQ induced Lcn2 –/– mice = 4. Representative images are shown (Bar = 200 μm)

Article Snippet: HFKs were cultured on coverslips and stimulated with either rmLCN2 or PBS, then fixed, permeabilized, and subjected to staining using TWEAK antibody sourced from Novus Biologicals (CO, USA).

Techniques: Immunostaining, Control, Staining, Software, RNA Sequencing, Knock-Out

Journal: Cellular and Molecular Immunology

Article Title: Synergistic effects of LCN2 and TWEAK on the progression of psoriasis

doi: 10.1038/s41423-025-01292-9

Figure Lengend Snippet: TWEAK promotes neutrophil expression of LCN2 but enhances inflammation in keratinocytes by inducing MMP9 expression. A Relative mRNA expression level of Lcn2, Tnfrsf12a , Tgfb1, Il6, Tnfa, Il1b, Krt5, Krt10, Krt17, 24p3r, Mc4r in mouse skin treated with TWEAK was detected by RT-qPCR. B Immunostaining of LCN2 and Ly6G were performed on paraffin sections of IMQ mice model with or without dorsally topical application of rmTWEAK (20 μg/mL, prepared in PBS). Unpaired two-tailed t test. C Immunofluorescence assays were used to detect LCN2 expression in neutrophils after TWEAK stimulation. Bar = 20 μm. D RT-qPCR was conducted to detect Lcn2 mRNA level in neutrophils treated with TWEAK. E LCN2 levels in neutrophil supernatants after TWEAK stimulation for 24 h were measured by ELISA. F Proteomic analysis was used to further investigate the protein changes and possible pathways following TWEAK action on keratinocytes. G After treating with TWEAK on primary keratinocyte with or without M5 cytokines for 48 h, LCN2 and 24P3R protein levels in keratinocytes were detected by Western blotting. H After the TWEAK stimulus, proteins such as MMP9 and TRAF2 were detected by Western blotting. Data from immunofluorescence assays and ELISA assay are representative of three or more independent experiments are presented as the means ± SEM of triplicate wells. ANOVA was used for comparison between groups: * P < 0.05, ** P < 0.01, and *** P < 0.001. ns not significant

Article Snippet: HFKs were cultured on coverslips and stimulated with either rmLCN2 or PBS, then fixed, permeabilized, and subjected to staining using TWEAK antibody sourced from Novus Biologicals (CO, USA).

Techniques: Expressing, Quantitative RT-PCR, Immunostaining, Two Tailed Test, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Western Blot, Comparison