Journal: bioRxiv
Article Title: Prolactin links psychological stress to psoriasis via a fibroblast-chemokine pathway
doi: 10.1101/2025.10.11.681540
Figure Lengend Snippet: a Schematic representation of the cell culture. b Volcano plot of differentially expressed genes between PBS-and PRL-stimulated NIH-3T3 cells ( n = 3). c KEGG pathway enrichment analysis of differentially expressed genes. d GSEA gene enrichment analysis of the chemokine signaling pathway. e Heatmap of chemokines. f The expression of Ccl2, Ccl7, Ccl8 , and Ccl11 as determined by quantitative PCR ( n = 3). g The expression of Ccl2 and Ccl7 in the skin of IMQ-treated female and male mice ( n = 3–4). h GSEA gene enrichment analysis of the JAK-STAT signaling pathway. i Western blot analyses of p-JAK2, JAK2, p-STAT1, STAT1, p-STAT3, STAT3, p-STAT5, and STAT5 in PRL-stimulated NIH-3T3 cells. j Western blot analyses of p-JAK2, JAK2, p-STAT3, and STAT3 as indicated. k The expression of Ccl2 and Ccl7 in PBS-, PRL+Veh-, and PRL+J2S3i-treated NIH-3T3 cells ( n = 3). Data are representative of two ( f, g, i – k ) independent experiments. Data are mean ± SD ( f, g, k ). The P values were calculated by two-tailed unpaired Student’s t -test ( f, g ) or one-way ANOVA with Dunnett’s post-hoc test ( k ); * P < 0.05, ** P < 0.01.
Article Snippet: For RNA-seq, the cells were stimulated with recombinant human PRL (100 ng/mL, MCE, #HY-P71059) for 6 h. For Western blot, the cells were stimulated with different concentrations of PRL (0, 1, 10, and 100 ng/mL) for 30 min, or transfected with siRNA targeting Prlr (5’-UAAAGAAACAGGAAUUGGGTT-3’) or scrambled control for 24 h and then stimulated with PRL (100 ng/mL) for 30 min. For quantitative PCR, the cells were pretreated with JAK2/STAT3 inhibitor FLLL32 (10 μM) for 1 h followed by stimulation with PRL (100 ng/mL) for 6 h.
Techniques: Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Two Tailed Test