ifit1 antibody  (Cell Signaling Technology Inc)

Journal: Journal of Human Immunity

Article Title: Human ISG15 deficiency unveils impaired healing of ulcerations via type I interferon–mediated fibrosis

doi: 10.70962/jhi.20250011

Figure Lengend Snippet: Identification of a patient with a novel ISG15 mutation. (A) Family includes two affected children carrying a homozygous variant (c.463insC) on ISG15 and six children of unknown status. The arrow indicates the patient described in this section (proband). (B) CT of the younger sibling taken at age 12. Extensive diffuse ground-glass opacities with subpleural sparing and enlargement of the main pulmonary artery are observed. (C) Schematic localization of the ISG15 variant in the genomic DNA locus indicated by the arrow. (D) Expression levels of various ISGs ( IFI27 , MX1 , SIGLEC1 , and IFIT1 ) were assessed from patient’s whole blood, as well as from the three healthy controls. Target genes were normalized on 18S rRNA expression. (E) 3′-RACE PCR targeting ISG15 was performed on RNA isolated from patient’s whole blood. (F) Sanger sequencing of the 3′-RACE band from E. (G) HEK293T cells were transfected with a plasmid encoding the ISG15 variant (c.463insC), ISG15 WT, or Luc. Relative mRNA levels for ISG15 were assessed by qRT-PCR, performed twice for each variant, with technical triplicates; the data from one representative experiment ( n = 3) are shown. The expression level of ISG15 was normalized to GAPDH . (H) HEK293T cells were transfected with a plasmid encoding the ISG15 variant (c.463insC), ISG15 WT or Luc, plus HERC5, UBE1L, and UBCH8, as described in . Cell lysates were analyzed by western blotting for ISG15 and ISGylation; a representative experiment is shown. CT, chest tomography. Source data are available for this figure: .

Article Snippet: The probes used were the following: TGFB1 Hs07289533_m1, ISG15 Hs01921425_s1, IFIT1 Hs03027069_s1, IFI27 Hs01086373_g1, SIGLEC1 Hs00988063_m1, MX1 Hs00895608_m1, FGF-2 Hs00266645_m1, ITGB6 Hs00168458_m1, PDGFA Hs00234994_m1.

Techniques: Mutagenesis, Variant Assay, Expressing, Isolation, Sequencing, Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Tomography

Journal: Journal of Human Immunity

Article Title: Human ISG15 deficiency unveils impaired healing of ulcerations via type I interferon–mediated fibrosis

doi: 10.70962/jhi.20250011

Figure Lengend Snippet: IFN-I inflammation in patient skin biopsies. (A) DEGs for each cluster were used to manually annotate the different clusters and then spatially visualized with one representative control (top). Percent composition of each cluster was calculated per biopsy (bottom). (B) Percentage of spots out of total biopsy that express each of the mentioned ISGs. (C) Localized expression and intensity levels of IFI44L . (D) Spatial expression of various ISGs. Red spots express at least one of the following ISGs: IFIT1 , USP18 , MX1 , IFI27 , IFI44L , or OAS1 . White spots represent spots that do not express any of those ISGs. (E) Percentages of spots out of total biopsy, epidermis only, and dermis only that express IFNB1 . P values were calculated with two-tailed t test. *P < 0.05; **P < 0.01.

Article Snippet: The probes used were the following: TGFB1 Hs07289533_m1, ISG15 Hs01921425_s1, IFIT1 Hs03027069_s1, IFI27 Hs01086373_g1, SIGLEC1 Hs00988063_m1, MX1 Hs00895608_m1, FGF-2 Hs00266645_m1, ITGB6 Hs00168458_m1, PDGFA Hs00234994_m1.

Techniques: Control, Expressing, Two Tailed Test

Journal: Journal of Human Immunity

Article Title: Human ISG15 deficiency unveils impaired healing of ulcerations via type I interferon–mediated fibrosis

doi: 10.70962/jhi.20250011

Figure Lengend Snippet: Cell death markers identified through ST. (A) Left: percentage of spots expressing apoptosis markers ( CASP3 , CASP8 , BAX , BAK1 , and CYCS ) out of total biopsy. Middle and right: percentage of spots expressing either two or five apoptosis markers in total biopsy, epidermis only, or dermis only (respectively). (B) Left: percentage of spots expressing necroptosis markers ( RIPK1 and MLKL ) out of total biopsy. Middle and right: percentage of spots expressing either RIPK1 or MLKL in total biopsy, epidermis only, or dermis only (respectively). (C) Percentage of spots expressing at least one apoptotic marker ( CASP3 , CASP8 , BAX , BAK1 , or CYCS ) that also express or not at least one of the ISGs ( IFIT1 , USP18 , MX1 , IFI27 , IFI44L , or OAS1 ). (D) Percentage of spots out of total biopsy, epidermis only, and dermis only that express ZBP1 . P values were calculated with a two-tailed t test. *P < 0.05; **P < 0.01.

Article Snippet: The probes used were the following: TGFB1 Hs07289533_m1, ISG15 Hs01921425_s1, IFIT1 Hs03027069_s1, IFI27 Hs01086373_g1, SIGLEC1 Hs00988063_m1, MX1 Hs00895608_m1, FGF-2 Hs00266645_m1, ITGB6 Hs00168458_m1, PDGFA Hs00234994_m1.

Techniques: Expressing, Marker, Two Tailed Test

Journal: iScience

Article Title: Single-cell transcriptomic analysis reveals intra-tumoral heterogeneity and immunotherapy strategies in high-grade serous ovarian cancer

doi: 10.1016/j.isci.2026.115266

Figure Lengend Snippet: Validation of tumor-associated marker genes (A) Expression of tumor-associated markers in epithelial cells from different cancers. (B) Scoring for major cell types based on tumor-associated markers. (C) Scoring of tumor diagnostic marker-related genes for major cell types. (D) Scoring of EV marker genes for major cell types. (E) Expression levels of co-stimulatory molecules and MHC molecules in different epithelial cell clusters. (F) Expression levels of top 2 genes in different epithelial cell clusters. (G and H) mIHC images displaying the expression of IFIT1 , epithelial marker (pan-CK), MHC class Ⅰ marker (B2M), and CD8 + T cell markers (CD3 + /CD8 + ) in HGSOC patients with (G) unfavorable prognosis and (H) favorable prognosis; n = 50. (I–K) CD8 + T cells (effectors) were incubated with IFIT1 + or IFIT1 − tumor cells (targets) at a 10:1 effector-to-target (E:T) ratio for 72 h. (I) Schematic of the in vitro culture model. (J) Representative flow cytometry plots of the results of the apoptosis assays and the quantification of apoptosis assays based on their Annexin V + percentages. (K) Levels of IFN-γ and granzyme B in the supernatant, measured via ELISA. Data are presented as the mean ± SEM, n = 5 pairs, including 5 patients and 5 healthy volunteers (J and K). ∗ p < 0.05, ∗∗ p < 0.01, two-tailed Student’s t test.

Article Snippet: IFIT1 antibody , Cell Signaling Technology , Cat# 14769 RRID: AB_2783869.

Techniques: Biomarker Discovery, Marker, Expressing, Diagnostic Assay, Incubation, In Vitro, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test