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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , <t>FGF2</t> ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.
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Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.

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: Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.

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: Activation Assay, Expressing, Generated, Two Tailed Test