Journal: Nature Communications

Article Title: Critical role of interferons in gastrointestinal injury repair

doi: 10.1038/s41467-021-22928-0

Figure Lengend Snippet: a 6–8-week-old naive WT, single or double IFNR-deficient mice receiving regular water were euthanized and proliferation of colon epithelial cells were evaluated by immunohistochemistry (IHC) staining for Ki-67 ( n = 5). b 6–8-week-old WT, single or double IFNR-deficient mice were exposed to 1.5% DSS in drinking water for 7 days, followed by regular drinking water for recovery. Mice were euthanized on day 8 and proliferation of colon epithelial cells was evaluated by IHC staining for Ki-67 ( n = 5). Representative colon images and quantified data are shown. Scale bars are 100 μM. Data are representative of two independent experiments. Symbols represent the number of Ki-67 + cells per crypt (five crypt/slide/animal; n = 5). Quantitative data were analyzed using one-way ANOVA followed by Bonferroni’s multiple comparisons test and represent mean values with SEM. **** P ≤ 0.0001 and are for the indicated animal group compared with control group of WT mice.

Article Snippet: For qPCR analysis, mIECs (mIEC-1 clone G7, mIEC-G7), murine lung epithelial cells (MLE-15, ATCC), and FAC-sorted CD45 + cells were treated for 3 h with 100 ng/ml of respective IFN (recombinant mouse IFN-α2, Novoprotein; recombinant mouse IFN-λ2, either from Peprotech or Bristol-Myers Squibb (pegylated or nonpegylated forms)).

Techniques: Immunohistochemistry, Control

Journal: Nature Communications

Article Title: Critical role of interferons in gastrointestinal injury repair

doi: 10.1038/s41467-021-22928-0

Figure Lengend Snippet: a–c Indicated bone marrow chimera mice and age-matched WT and Ifnar1 −/− Ifnlr1 −/− mice were exposed to 1.5 % DSS in drinking water for 7 days, followed by regular water for recovery. Mice were monitored daily for a survival and b weight loss (WT, n = 8; WT→WT, n = 5; Ifnar1 −/− Ifnlr1 −/− , n = 14; Ifnar1 −/− Ifnlr1 −/− → Ifnar1 −/− Ifnlr1 −/− , n = 4, Ifnar1 −/− Ifnlr1 −/− →WT, n = 10; WT→ Ifnar1 −/− Ifnlr1 −/− , n = 10; Ifnar1 −/− → Ifnlr1 −/− , n = 9). Data are presented as mean values with SEM. c Mice were euthanized on day 8 and colon epithelial cell proliferation was evaluated by IHC staining for Ki-67 ( n = 5). d 6–8-week-old WT mice were treated with IFN (1.0 μg) or PBS as a control by subcutaneous injection, and euthanized 30 min later. IFN-induced activation of STAT1 was evaluated by IHC staining with antibodies against phosphorylated STAT1 ( n = 5). Scale bars are 100 μM. Data are representative of two independent experiments ( a–d ).

Article Snippet: For qPCR analysis, mIECs (mIEC-1 clone G7, mIEC-G7), murine lung epithelial cells (MLE-15, ATCC), and FAC-sorted CD45 + cells were treated for 3 h with 100 ng/ml of respective IFN (recombinant mouse IFN-α2, Novoprotein; recombinant mouse IFN-λ2, either from Peprotech or Bristol-Myers Squibb (pegylated or nonpegylated forms)).

Techniques: Immunohistochemistry, Control, Injection, Activation Assay

Journal: Nature Communications

Article Title: Critical role of interferons in gastrointestinal injury repair

doi: 10.1038/s41467-021-22928-0

Figure Lengend Snippet: a, b Mx2-Luciferase reporter mice ( n = 5) were exposed to 1.5% DSS in drinking water for 7 days, followed by regular water. Mice were injected with luciferin and levels of IFN-controlled Mx2 promoter-driven luciferase expression were measured by IVIS on selected days. c, d 6–8-week-old WT and Ifnar1 −/− Ifnlr1 −/− mice were administered 1.5% DSS in drinking water for 7 days, followed by regular water. Levels of colonic AREG transcripts were analyzed on day 11 by c in situ hybridization ( n = 5) and d qPCR from total colon homogenates ( n = 5). e Murine intestinal epithelial cells (mIECs) or f fluorescent-activated cell (FAC)-sorted bone marrow-derived CD45 + cells were treated with IFN for 30 min and upregulation of AREG transcripts were analyzed by qPCR ( e , n = 6; f , n = 5). Scale bars are 100 μM. Data are representative of two ( a, b ) or three independent experiments ( c ) or pooled from two independent experiments ( d–f ). Symbols represent individual data points ( d–f ). Quantitative data were analyzed using b, e, f one-way ANOVA or d two-tailed unpaired t -test followed by Tukey’s ( b ) or Bonferroni’s multiple comparisons test ( e, f ) and represent mean values with SEM. b * P = 0.0231 (day 13), * P = 0.0168 (day 22), ** P = 0.0055 (day 18), *** P = 0.0004 (day 8), *** P = 0.0005 (day 19); P values are for the indicated day compared with day 0. d **** P ≤ 0.0001 and is for the indicated animal group compared with control group of WT mice. e ** P = 0.0027, **** P ≤ 0.0001. f * P = 0.0483 (IFN-λ), ** P = 0.0023 (IFN-α), ** P = 0.005 (IFN-α/IFN-λ). P values are for the indicated treatment compared with mock ( e, f ).

Article Snippet: For qPCR analysis, mIECs (mIEC-1 clone G7, mIEC-G7), murine lung epithelial cells (MLE-15, ATCC), and FAC-sorted CD45 + cells were treated for 3 h with 100 ng/ml of respective IFN (recombinant mouse IFN-α2, Novoprotein; recombinant mouse IFN-λ2, either from Peprotech or Bristol-Myers Squibb (pegylated or nonpegylated forms)).

Techniques: Luciferase, Injection, Expressing, In Situ Hybridization, Derivative Assay, Two Tailed Test, Control

Journal: Nature Communications

Article Title: Critical role of interferons in gastrointestinal injury repair

doi: 10.1038/s41467-021-22928-0

Figure Lengend Snippet: a–c 6–8-week-old WT and Ifnar1 −/− Ifnlr1 −/− were exposed to 1.5% DSS in drinking water for 7 days, followed by regular water. On days 5, 7, 9, and 11, Ifnar1 −/− Ifnlr1 −/− mice were administered AREG (5.0 μg) or PBS as a control by IP injection, and the animals were monitored daily for survival ( a ) ( n = 5). b , c On day 11, mice were euthanized ( n = 5) and proliferation of epithelial cells in the colon was evaluated by IHC staining for Ki-67. Symbols in b represent the number of Ki-67 + cells per crypt (five crypt/slide/animal; n = 5). c Representative images. Scale bars are 100 μM. Data are representative of two independent experiments. Quantitative data were analyzed using one-way ANOVA followed by Bonferroni’s multiple comparisons test and represent mean values with SEM. **** P ≤ 0.0001.

Article Snippet: For qPCR analysis, mIECs (mIEC-1 clone G7, mIEC-G7), murine lung epithelial cells (MLE-15, ATCC), and FAC-sorted CD45 + cells were treated for 3 h with 100 ng/ml of respective IFN (recombinant mouse IFN-α2, Novoprotein; recombinant mouse IFN-λ2, either from Peprotech or Bristol-Myers Squibb (pegylated or nonpegylated forms)).

Techniques: Control, Injection, Immunohistochemistry