Journal: Journal of translational medicine
Article Title: Potential of CLSPN as a therapeutic target in melanoma: a key player in melanoma progression and tumor microenvironment.
doi: 10.1186/s12967-025-06455-w
Figure Lengend Snippet: Fig. 4 RNA sequencing of A2058 cells to investigate the function of CLSPN. (A) Volcano plot of the differential gene expression between CLSPN knock down group and the control group. The red points represented significantly up-regulated gene, and the blue points represented significantly down-reg ulated gene. (B) The top 20 enrichment of KEGG pathway terms analysis for the differential genes.(C) Protein-protein interaction analysis of differentially expressed functional genes. (D) Molecular docking result of CLSPN/IFI44L. (E) The mRNA expression of IFI44L and downstream genes after CLSPN knock down in A2058 by qPCR. (F) The protein levels of CLSPN, IFI44L, p-STAT1, STAT1 and XAF1 after si-CLSPN transfection by western blot. (G) The quantifica tion results of western blot by Image J (n = 3). Data were expressed as mean ± SD. * p < 0.05 and ** p < 0.01
Article Snippet: The membranes were blocked using 5% non-fat milk for 1 h, incubated overnight with primary antibodies against CLSPN (Affinity, DF7525), IFI44L (SAB, 27948), p-STAT1 (Proteintech, 28977-1-AP), STAT1 (Proteintech, 10144-2-AP), XAF1 (BOSTER, BA4986), GAPDH (Affinity, AF7021), antiBax (Cell Signaling Technology, #5023), anti-Bcl-2 (Cell Signaling Technology, #3498), anti-cleaved-PARP (Cell Signaling Technology, #5625T), anti-cleaved-caspase 9 (Cell Signaling Technology, #20750), anti-β-tubulin (Affinity, AF7011), anti-β-actin (Affinity, #AF7018) at 4 °C, and then incubated for 1 h with secondary antibody goat Anti-Rabbit IgG (Affinity, #S0001).
Techniques: RNA Sequencing, Gene Expression, Knockdown, Control, Functional Assay, Expressing, Transfection, Western Blot