Fig 2: DSV-induced Notch signaling, SOCS3, and IL-1β expression is inhibited by Notch1siRNA (A) RAW cells were transfected with either scrambled (Scr) or siRNA against Notch1 (siNotch1) by electroporation. After incubation for 24 hours, cells were removed and re-plated at a density of 8x105 and further incubated for 24 hours. Following day (total of 48 hrs transfection), cells were infected with DSV (MOI 20) for 7 hours. Cells were lysed and protein lysate was prepared. Fifty μg of protein lysate was separated on SDS-PAGE and analyzed for Notch1, NICD, SOCS3, IL-1β, and p21 by Western blotting. Actin was used as a loading control. (B–F) Quantification of Western blots. Blots were quantified with ImageJ by analyzing the ratio of protein of interest/Actin. Data represents Mean ± SEM from at least three independent experiments. One-way ANOVA was used to determine the statistical significance. Values were compared to Scr+ DSV, with a post-hoc Dunnett’s test. *p < 0.05, **p < 0.01.

Fig 3: (A) GSEA analysis was employed to explore cancer hallmarks that enriched high and low SOCS3 expression. (B) GSEA showed that IL‐6/JAK/STAT3 signalling pathway was differentially enriched in SOCS3. (C,D) Western blot revealed the relationship between SOCS3 and JAK/STAT3 signalling pathway.

Fig 4: (A–D) The heat map showed the upregulated and downregulated top 10 differentially expressed genes (DEGs). Positive and negative DEGs of SOCS1 (A), SOCS2 (B), SOCS3 (C), SOCS4 (D) according to heat map and Venn diagram. (E) Intersection representing DEGs of SOCS1/2/3/4. |r| > 0.4 and p < 0.001. (F) Gene network associated with SOCS1/2/3/4 and interactions between proteins encoded by genes of SOCS1/2/3/4 using GeneMANIA. (G) Correlation analysis for SOCS1/2/3/4. (H) The total significantly associated genes with SOCS3 distinguished by Pearson test in the TCGA‐GBM cohort. (I,J) Top 50 positively and negatively associated with SOCS3 in TCGA‐GBM were shown, respectively, via heat maps. Red, positively linked genes; blue, negatively linked genes.

Fig 5: The ROC curve to test the value of SOCS1/2/3/4 to identify GBM tissues was constructed. (A) SOCS1; (B) SOCS2; (C) SOCS3; (D) SOCS4. The results were 0.852, 0.962, 0.864 and 0.914, respectively, which suggests that SOCS1/2/3/4 genes may have the potential to identify GBM samples. (E) Nomogram model predicting the 1‐, 3‐ and 5‐year overall survival in patients with GBM. The nomogram is employed by summing all points identified on the scale for each variable, including gender, race, age, IDH status, Karnofsky performance score and SOCS1/2/3/4 expression in GBM. The total points projected on the bottom scales reveal the probabilities of 1‐, 3‐and 5‐year overall survival.