il5 rfp cre mice  (Jackson Laboratory)

Journal: JCI Insight

Article Title: Vancomycin eliminates gut deoxycholic acid, restoring ER proteostasis in ILC2s and relieving colitis

doi: 10.1172/jci.insight.197470

Figure Lengend Snippet: ( A – C ) Large intestinal ILC2s were sorted from WT mice and cultured in the presence of IL-2, IL-7, IL-25, and IL-33 for 5 days. ILC2s were treated with DCA (100 μM) at the indicated concentrations for 24 hours and then subjected to RNA-Seq. ( A ) Experimental strategy, ( B ) volcano plot, and ( C ) heatmap of ILC2-characteristic genes. ( D – F ) Sorted large intestinal ILC2s from Il5 RFP-Cre mice were treated with DCA (100 μM) for 24 hours. ( D ) Experimental strategy. ( E ) Representative flow cytometry plots. ( F ) Percentages of RFP + ILC2s (CD45.2 + Lin – GATA3 + ) in large intestine. n = 6 wells per group. The experiment was repeated 3 times. ( G – I ) Rabbit reticulocyte lysate system. ( G ) Experimental strategy. Protein levels of Areg, IL-5, and IL-13 were analyzed by ( H ) Western blotting and ( I ) cytometric bead array in samples treated or untreated with DCA. n = 6 wells per group. The experiment was repeated twice. Data are shown as mean ± SD. Statistical analysis was performed using unpaired 2-tailed t test. *** P < 0.001, **** P < 0.0001.

Article Snippet: Il5 RFP-Cre mice (catalog R5/+) and CD45.1/CD45.1 mice (catalog 002014) were purchased from The Jackson Laboratory.

Techniques: Cell Culture, RNA Sequencing, Flow Cytometry, Western Blot

Journal: Advanced Science

Article Title: Sensing of Liver‐Derived Nicotinamide by Intestinal Group 2 Innate Lymphoid Cells Links Liver Cirrhosis and Ulcerative Colitis Susceptibility

doi: 10.1002/advs.202404274

Figure Lengend Snippet: NAMPT deficiency in ILC2s aggravates DSS‐induced colitis. A,B) Representative FACS analyses and absolute numbers of ILC2s in the large intestine of Nampt +/+ Il5 RFP‐Cre/+ or Nampt f/f Il5 RFP‐Cre/+ littermate mice. Data were presented as mean ± SD; n = 8 per group, from four independent experiments. C,D) Representative FACS analyses and frequency of RFP + ILC2s after gating on Lin – KLRG1 + ILC2s in the LI of Nampt +/+ Il5 RFP‐Cre/+ or Nampt f/f Il5 RFP‐Cre/+ littermate mice; mean ± SD of 8 biological replicates; from three independent experiments. E,F) Large intestinal ILC2s were sorted from Nampt +/+ Il5 RFP‐Cre/+ or Nampt f/f Il5 RFP‐Cre/+ littermate mice and cultured in the presence of IL‐2, IL‐7, IL‐25, and IL‐33 for 7 days. The nicotinamide mononucleotide (NMN) or succinate were added on day 0, and ILC2s were collected at day 7. Experimental strategy (E). The absolute numbers of ILC2s and frequency of Areg, IL‐5, and IL‐13 expression in ILC2s (CD45.2 + Lin – GATA3 + ) (F). Data were presented as mean ± SD; n = 6 per group, from three independent experiments. G,H) The lethally irradiated Rag2 –/– Il2rg –/– recipient mice were transferred with an equal mixture (1:1) of bone marrow cells from Nampt +/+ Il5 RFP‐Cre/+ (WT, CD45.1/CD45.2) or Nampt f/f Il5 RFP‐Cre/+ (cKO, CD45.2/CD45.2) mice. Experimental strategy (G). Representative FACS analyses and frequency of ILC2s, Areg + ILC2s, IL‐5 + ILC2s, and IL‐13 + ILC2s from indicated donors (H). Data were presented as mean ± SD; n = 5 per group, from two independent experiments. I–N) The Nampt f/f Il5 RFP‐Cre/+ mice were orally administrated NMN for two weeks before DSS administration. Representative colons (I) and colon lengths (J), representative hematoxylin‐eosin (H&E) staining (K) and histological severity scores of colons (L). Flow cytometry analysis of Lin – GATA3 + ILC2s expression (M), absolute numbers of ILC2s, Areg + , IL‐5 + and IL‐13 + ILC2s (N); mean ± SD of 7 biological replicates, from two independent experiments. The two‐sided unpaired t‐test in B, D, H, J, L, N. The One‐way ANOVA with Dunnett in F. The statistical test method and exact P value are detailed in the Table (Supporting Information). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Il5 RFP‐Cre mice and CD45.1/CD45.1 mice were purchased from Jackson Laboratories.

Techniques: Cell Culture, Expressing, Irradiation, Staining, Flow Cytometry