Fig 1: IFIT2 staining in human ccRCC tissues and normal tissues. Positive immunostaining of IFIT2 could be detected in the cytoplasm of cancer cells in ccRCC tissues or of renal tubular epithelial cells in normal tissues. IFIT2, interferon (IFN)-induced protein with tetratricopeptide repeats 2; ccRCC, clear cell renal cell carcinoma.
Fig 2: Increases in host cellular mRNA does not correlate with increases in protein expression. (a) HSAECs were mock-infected or infected with MP12 for one hour. At 18 hours post infection, cell lysates were collected and analyzed by western blot. As a control, U937 cell lysates were used. At 24 hours post infection, cell lysates were collected and analyzed in parallel by western blot. Membranes were probed for levels of ARHGEF6 (α-PIX) and β-Actin as a loading control. (b) HSAECs were mock-infected, infected with MP12, or infected with MP12-ΔNSs for one hour. Following infection, cells were treated with Actinomycin D (1 µg/mL) for the remaining time. At 18 hours post infection, cell lysates were collected and analyzed in parallel by western blot for levels of IFIT2, Viperin, α-Actinin-2, and β-Actin as a loading control. Cropped, representative images are shown here. Uncropped western blots can be found in Supplemental Fig. 8.
Fig 3: Immunohistochemical staining of IFIT2 in lung cancer tissue and adjacent normal tissue. (A) Wilcoxon rank test showed that H-score in lung adenocarcinoma tissues was significantly lower than that in adjacent normal tissue (P< 0.0001). (B) Wilcoxon rank sum test showed that H-score in lung squamous cell carcinoma was significantly lower than that in normal tissue (P< 0.0001). (C) The correlation between IFIT2 expression and OS in lung adenocarcinoma: Kaplan–Merier survival analysis showed that the OS of patients with low IFIT2 expression was significantly poorer than that of patients with high IFIT2 expression (HR = 2.392, 95% CI: 1.103–5.186, P = 0.027). (D) The correlation between IFIT2 expression and OS in lung squamous cell carcinoma: Kaplan–Merier survival analysis showed that the OS of patients with low IFIT2 expression was significantly poorer than that of patients with high IFIT2 expression (HR = 2.907, 95% CI: 1.118–7.559, P = 0.029).Abbreviation: IFIT2, interferon-induced proteins with tetratricopeptide repeats 2.
Fig 4: Analysis of differentially expressed genes in HSAECs following RVFV infection. (a) Venn diagrams depict the upregulated (top panel) and downregulated (bottom panel) genes changed in MP12 only (left), ZH548 only (right), or both (center, gray) at 3, 9, and 18 hours post infection. These genes were changed by 1.5-fold or more and had a p-value ≤ 0.05. (b) HSAECs were mock-infected or infected with MP12 or ZH548 at MOI 5 for one hour. Lysates were collected in Trizol LS, RNA was extracted and prepared for RNA sequencing. RNA-sequencing reads were normalized to the total reads, then fold changes were calculated against the uninfected, mock samples at the specified time point. (c) RT-qPCR confirmation of some of the top changed transcripts during all time points post infection. HSAECs were mock-infected or infected with MP12 or ZH548 at MOI 5 for one hour. Lysates were collected in Trizol LS, RNA was extracted using the Direct-zol™ RNA MiniPrep, and analyzed for RT-qPCR with TaqMan Gene Expression Assays against IFIT1, IFIT2, IFIT3, and RSAD2. Fold changes were calculated relative to 18 S ribosomal RNA and normalized to mock samples using the ΔΔCt method. Data are expressed as the Mean ± SD (n = 3).
Fig 5: Prognostic value of IFIT2 staining in ccRCC tissues. (A) The expression level of IFIT2 in renal cancer tissues was significantly lower than that in adjacent normal tissues (P<0.001); (B) higher IFIT2 expression in ccRCC tissues was significantly associated with better patient survival (HR: 0.428, 95% CI: 0.199–0.923, P=0.030); (C,D) based on TCGA gene profile data from http://gepia.cancer-pku.cn/, our results revealed that decreased IFIT2 mRNA expression in ccRCC tissues was significantly correlated with poor OS and DFS in patients (P=0.017 and P=0.11, respectively). IFIT2, interferon (IFN)-induced protein with tetratricopeptide repeats 2; ccRCC, clear cell renal cell carcinoma; TCGA, The Cancer Genome Atlas; OS, overall survival; DFS, disease-free survival.
Supplier Page from Abcam for Anti-IFIT2 antibody