rabbit anti irf1  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: Single-cell transcriptomics reveals aberrant skin-resident cell populations and identifies fibroblasts as a determinant in rosacea

doi: 10.1038/s41467-024-52946-7

Figure Lengend Snippet: a UMAP plots showing keratinocyte subclusters and sample conditions. b Bar graph showing subpopulation percentage. c Feature plot showing CD74 expression in keratinocytes. d The percentage of CD74 + keratinocytes ( n = 3 samples for each group). e Immunohistochemistry of KRT14, CD74 in HS, normal skin of patients (NS), and lesional skin of ETR, PPR, PhR. White arrows indicate CD74 + keratinocytes. Scale bar, 50 μm. f Quantification of CD74 + keratinocytes ( n = 6/6/6/6/5 samples for HS/NS/ETR/PPR/PhR group used in e ). g , h Top-ranked enriched pathways in CD74 + keratinocytes of L versus HS ( g ), L versus N ( h ). NES indicates enrichment scores. The color keys from red to blue indicate the P -value range. Two-sided permutation test without multiple comparison adjustments was used. i Heatmap of skin barrier-related genes in CD74 + keratinocytes. j Heatmap of IFN-related genes in CD74 + keratinocytes. k Immunohistochemistry of IRF1, KRT14. Scale bar, 100 μm. l Quantification of IRF1 + keratinocytes in epidermis n = 6/6/6/6/5 samples for HS/NS/ETR/PPR/PhR group used in k ). m The back skins of IgG and anti-IFNγ antibody-treated mice intradermally injected with LL37 or control vehicle. Images were acquired 48 h after the first LL37 injection. Below panels, magnified images of yellow boxed areas. n The severity of the rosacea-like symptoms was determined with the redness area and score ( n = 6). o Dermal infiltrating cells were quantified ( n = 6). p The relative mRNA levels of Il1β, Il6 , Tnfα in mice skins ( n = 6). q The relative mRNA levels of Cldn4 , Cldn10 , Cldn23 in mice skins ( n = 6). r Immunohistochemistry of CLDN4, KRT14 in mice skins. Scale bar, 50 μm. Epi, Epidermis. Der, Dermis. s Quantification of relative fluorescence intensity for CLDN4 in epidermis ( n = 6). t Quantification of TEWL ( n = 6). All results are representative of at least three independent experiments. Data are presented as mean ± SEM, and P -values were determined by one-way ANOVA with Tukey’s post hoc test ( f , l , n , o , p , q , s , t ) and two-tailed unpaired (L vs HS) or paired (L vs N) Student’s t -test ( d ).

Article Snippet: Rabbit anti-p-STAT1 (1:1000, 9167, Cell signaling), Rabbit anti-IRF1 (1:200, 8478, Cell signaling).

Techniques: Expressing, Immunohistochemistry, Comparison, Injection, Control, Fluorescence, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: IRF2 loss is associated with reduced MHC I pathway transcripts in subsets of most human cancers and causes resistance to checkpoint immunotherapy in human and mouse melanomas

doi: 10.1186/s13046-024-03187-5

Figure Lengend Snippet: Transcription factor IRF1 substitutes for the loss of IRF2. A Same experimental design as 5C and 5D except, IRF1 mRNA expression was analyzed by qPCR. B-E EV, IRF1KO, IRF2KO and DKO (IRF1+IRF2KO) B16F0 cells were stimulated with IFNα in vitro and analyzed by: B Western blot for IRF1 or ß-actin; C . qPCR expression for MHC I pathway components ( n= 2/group). D & E Surface MHC I levels after 0 or 100 ng/ml IFNα treatment for 24 h ( n= 2/group)

Article Snippet: After transfer, PVDF membranes (Millipore) were blocked with TBS-Tween 1x + 5% milk and then blotted with rabbit anti-IRF2 antibody (Abcam ab124744) or rabbit anti-IRF1 antibody (Abcam ab186384) in TBS-Tween 1x + 2% milk overnight at 4°C.

Techniques: Expressing, In Vitro, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: IRF2 loss is associated with reduced MHC I pathway transcripts in subsets of most human cancers and causes resistance to checkpoint immunotherapy in human and mouse melanomas

doi: 10.1186/s13046-024-03187-5

Figure Lengend Snippet: B16 melanoma IRF1+IRF2 double KO cells show impaired responses to IFN inducer Poly(I:C) plus CPI. A NSG mice ( n= 10) and C57BL/6 mice ( n= 12) were subcutaneously injected with WT, IRF2KO and IRF1+IRF1KO cells and data display tumor growth in individual mice. B Tumors were collected on day 15 and the surface MHC I expression of C57BL/6 tumors was analyzed by flow cytometry. C C57BL/6 mice ( n= 45), that were treated with poly(I:C) and aPD1 were injected with IFNα treated (10ng/mL, 24h) control WT vs IRF2KO IRF2+IRF1 (double) KO B16 cells and tumor growth was followed. D & E On day 17 post tumor injection, n= 5 mice/group were sacrificed for: D mRNA expression analysis of MHC I pathway components using qPCR method and E the tumor cell surface expression analysis of MHC I molecules by flow cytometer. Each dot represents the average measurement of the individual tumors collected from mice. F Survival analysis of WT or IRF2KO or IRF1+IRF2 KO tumors in C57BL/6 mice. This experiment was repeated twice. Tumor growth curve shows mean+SD ( C )

Article Snippet: After transfer, PVDF membranes (Millipore) were blocked with TBS-Tween 1x + 5% milk and then blotted with rabbit anti-IRF2 antibody (Abcam ab124744) or rabbit anti-IRF1 antibody (Abcam ab186384) in TBS-Tween 1x + 2% milk overnight at 4°C.

Techniques: Injection, Expressing, Flow Cytometry, Control