Journal: The Journal of Experimental Medicine

Article Title: RNase T2 deficiency promotes TLR13-dependent replenishment of tissue-protective Kupffer cells

doi: 10.1084/jem.20230647

Figure Lengend Snippet: Rnaset2 −/− mice are resistant to acute liver injuries. (A) Survival curve of indicated mice with APAP challenge at 750 mg/kg ( n = 11–15). (B) H&E staining of the liver of indicated mice 24 h after APAP challenge at 500 mg/kg. Scale bar, 400 µm. (C, D, F, and G) Dot plots show serum levels of AST (C), ALT (D), CXCL2 (F), and IL-6 (G) in indicated mice at 6 and 24 h after APAP challenge at 500 mg/kg ( n = 4–6). (E) Dot plots show the percentages of neutrophils that infiltrated the liver in indicated mice 24 h after APAP challenge at 500 mg/kg ( n = 4). (H) Percentage of TNF-α + cells in indicated hepatic macrophage subsets from wild-type and Rt2 −/− mice after in vitro culture with the TLR9 ligand CpG-B (200 nM) or the TLR4/MD-2 ligand lipid A (1 µg/ml) together with Brefeldin A for 3 h ( n = 3). (I) Percentage of TNFα + cells in hepatic Ly6C low macrophages in wild-type mice after in vitro culture with lipid A (1 µg/ml) and Brefeldin A for 3 h, with or without LXR agonist ( n = 3). (J) Survival curve of Rt2 −/− mice challenged with APAP at 750 mg/kg. Indicated mice had been intravenously administered with clodronate at 25 mg/kg 24 h before APAP challenge ( n > 4). (K) Survival curve of wild-type mice left untreated or administered twice with recombinant AIM (400 µg) before APAP challenge ( n = 10–15). (L) Survival curve of wild-type mice left untreated or administered twice with LXR agonist before APAP challenge ( n = 5–10). (M) Serum levels of AST and ALT in Rt2 −/− mice with or without antibiotic treatment ( n = 5). (N) Survival curve of wild-type mice left untreated or administered twice with the TLR13 ligand Sa19 before APAP challenge ( n = 5–8). *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: Phycoerythrin (PE)-conjugated mouse anti-TLR3 mAb (PaT3), PE-conjugated mouse anti-TLR7 mAb (A94B10), and PE-conjugated mouse anti-TLR9 mAb (J15A7) were purchased from BD Biosciences.

Techniques: Staining, In Vitro, Recombinant

Journal: Nature Communications

Article Title: An essential role for TASL in mouse autoimmune pathogenesis and Toll-like receptor signaling

doi: 10.1038/s41467-024-55690-0

Figure Lengend Snippet: a FACS histogram overlays of TLR7 or TLR9 expression from WT and Tasl –/– (KO) mice splenic pDCs. Solid lines indicate staining with primary antibodies, while dotted lines indicate antibody isotype staining controls. b Immunoprecipitation (IP) of TLR7 was performed on Flt3L pDCs derived from WT or Tasl –/– (KO) mice. Western blot analysis was performed for detection of TLR7 processing. HEK293T cells stably expressing mouse TLR7 and TLR4 was used respectively as a positive and negative control. β-actin in whole cell lysate (WCL) was detected as loading control. NS indicates a non-specific band. Data are representative of at least 3 independent experiments c Flt3L pDCs were stimulated with CpG A (3 μM) for the indicated time points. Western blot analysis and quantification was performed for JNK and total JNK and ( d ) p38 and p-p38. e Western blot analysis and quantification of IRF5 and p-IRF5 in CpG A (3 μM) stimulated Flt3L pDCs at the indicated time points. f Western blot analysis and quantification of p-IRF5 and IRF5 in in CpG A (3μM) stimulated splenic B cells at the indicated time points. Quantification of western blot is represented as the ratio of intensities relative to time zero. n = 5 mice per genotype were used and pooled per experiment.

Article Snippet: Anti-mouse TLR9 TIR domain serum , and anti-TLR7 N terminus (Invitrogen;PA112829) were used to detect TLR9 and TLR7 respectively.

Techniques: Expressing, Staining, Immunoprecipitation, Derivative Assay, Western Blot, Stable Transfection, Negative Control, Control