Journal: STAR Protocols

Article Title: Protocol for condensate-based stabilization of gene circuit dynamics under growth-mediated dilution in E. coli

doi: 10.1016/j.xpro.2026.104536

Figure Lengend Snippet: Design of gene circuits used in this protocol (A) DNA sequence of B0034-MCS1-pSB1A2 between RFC10 prefix and suffix. The solid green arrow marks the start codon of the open reading frame. (B) Schematic diagrams of synthetic gene circuits constructed and characterized in this study. Self-activation circuits include non–phase-separating controls (OP152 and OP177) and phase-separating circuits incorporating intrinsically disordered regions (OP153 and OP203). (C) Colocalization circuits include CT149, which contains 12× lacO operator sites adjacent to the target promoter, and IC41, a matched control lacking lacO sites. All circuits are based on AraC-mediated positive feedback and are expressed in E. coli.

Article Snippet: BglII-RFP- RFC10 surfix , E1010 , RFP-BglII-F , VR , 880 bp.

Techniques: Sequencing, Construct, Activation Assay, Control

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: bioRxiv

Article Title: Meningeal neutrophil infiltration drives inflammation-exacerbated pediatric stroke through IL-36γ signaling

doi: 10.64898/2026.04.03.716365

Figure Lengend Snippet: a, Expression of neutrophil activation markers (ICAM1, iNOS, CD62L and CXCR2) in the ipsilateral hemisphere was assessed by flow cytometry at 24 h post-stroke in PT and LPS/PT groups (N = 7 per group). Data were presented as mean ± S.E.M. ****p < 0.0001 compared with the PT group by unpaired t-test. b-c, Mice were treated with a neutrophil-depleting anti-Ly6G antibody (clone 1A8) or an isotype control antibody beginning one day prior to stroke induction and continuing daily for three consecutive days (P15–P17). Brain sections (1mm sections) were collected 3 days post-stroke and stained with TTC. Infarct volumes were quantified in mm3. (N> 10 per group). *p < 0.05 by one-way ANOVA. d, CCR2–RFP knock-in mice, including homozygous CCR2-deficient (CCR2RFP/RFP) and heterozygous CCR2-sufficient (CCR2RFP/+) littermates, were subjected to PT or LPS/PT. Brain sections were harvested 3 days post-stroke and infarct volumes were quantified by TTC staining and expressed as a percentage of the contralateral hemisphere (N > 5 per group). ns, Not significant, p>0.05; **p < 0.01; ***p < 0.001 by one-way ANOVA.

Article Snippet: C57BL/6J, CCR2 RFP/RFP (JAX#017586), and GFAP-Cre (JAX#012886) mice were purchased from The Jackson Laboratory.

Techniques: Expressing, Activation Assay, Flow Cytometry, Control, Staining, Knock-In