rat anti mouse clec4f  (R&D Systems)

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: ATG16L1 Regulates Reparative Function of Peritoneal Macrophages During Acute Drug-induced Liver Injury

doi: 10.1016/j.jcmgh.2025.101674

Figure Lengend Snippet: Depletion of PMs exacerbates APAP-induced liver injury in the early phase. Mice with depleted PMs and control mice were treated with APAP using the standard protocol. Liver and serum samples were collected at 6 and 12 hours thereafter. Representative H&E ( A ) staining, serum ALT ( B ) and AST ( C ), IL-1β ( D–E ), and IL-10 ( F–G ) levels were analyzed. Following intraperitoneal injection of control or CLL (100 μL), livers from WT mice were collected at day 7 post-injection. Tissue sections underwent dual-color IF staining for F4/80 and Clec4f, with the proportion of F4/80 + Clec4F + cells quantified by confocal microscopy analysis ( H ). n = 6/group. Data are shown as mean ± SEM. Each point represents an independent experiment. ∗ P < .05, ∗∗ P < .01.

Article Snippet: For immunohistochemical (IHC) and immunofluorescence (IF) staining, the sections were dehydrated and antigens were retrieved by incubation overnight at 4°C with the following primary antibodies: rat anti-mouse F4/80 (1:100, ab11101, Abcam), mouse anti-mouse GATA6 (1:200, 67943-1-Ig, proteintech), rabbit anti-mouse ATG16L1 (1:200, ab188642, Abcam), rabbit anti-mouse platelet endothelial cell adhesion molecule-1 (CD31) (1:100, ab222783, Abcam), hyaluronan-binding protein 1/complement component 1, q subcomponent binding protein (HABP1/C1QBP) (HY- P76185 , MCE), Mouse (His) (1:100, ab213204, Abcam), rabbit anti-mouse LC3 (1:200, ab192890, Abcam), rat anti-mouse CD11b (1:50, ab8878, Abcam), rat anti-mouse Ly6G (1:500, 88876, CST), rat anti-mouse Clec4f (MAB2784; 1/1000, R&D Systems), and rabbit anti-mouse PCNA (1:200, ab29, Abcam) antibodies.

Techniques: Control, Staining, Injection, Confocal Microscopy

Journal: Acta Pharmaceutica Sinica. B

Article Title: Palmitoylation of Tfr1 enhances platelet ferroptosis and liver injury in heat stroke

doi: 10.1016/j.apsb.2025.10.027

Figure Lengend Snippet: Platelet-derived IL-1 β contributes to liver injury in the HS murine model. (A) GO (BP) analysis of the physiological and pathological states of platelets in SHR mice and mice at 0 h post-HS (HS0). (B) Representative images of immunofluorescence in liver tissue from SHR and HS mice ( n = 3 independent experiments, scale bars, 50 μm). DAPI (blue), CD42b for platelets (green), Clec4f for Kupffer cells (red), Glutamine Synthetase (GS) for central vein zone (purple). (C) Flow cytometric analysis of the percentage of P-selectin expression on platelets in SHR mice and mice at 0, 6, 24 h post-HS ( n = 6 mice per group). (D) GO (BP) analysis of cytokine-related pathways in platelets from SHR mice and at HS onset (HS0). (E, F) qPCR analysis of Il1b (E) and Casp1 (F) mRNA expression of platelets under heat stress in vitro ( n = 6 mice per group). (G) Flow cytometry analysis of caspase-1 levels in platelets from Tfr1 flox/flox mice, CKO- Tfr1 mice and CKO- Tfr1 mice infected with AAV9-DIO- Tfr1 , with or without ferroptosis inhibitor fer1 and DFO ( n = 6 mice per group). (H) ELISA analysis of IL-1 β levels in the supernatant of platelets under heat stress in vitro ( n = 6 mice per group). (I) ELISA analysis of IL-1 β levels in plasma from Tfr1 flox/flox mice, CKO- Tfr1 mice, and CKO- Tfr1 mice infected with AAV9-DIO- Tfr1 , with or without ferroptosis inhibitor fer1 and DFO ( n = 6 mice per group). (J) ELISA analysis of IL-1 β levels in PLP supernatant from sh- Zdhhc2, oe- Zdhhc2 , oe- Tfr1 , oe- Tfr1 -sh- Zdhhc2 MEG-01 cells under heat stress in vitro , with or without ferroptosis inhibitor fer1 ( n = 6 independent experiments). Mean ± SEM are shown, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: After blocking, Clec4f (AF2784, R&D Systems, Minneapolis, MN, USA), Glutamine Synthetase (ab73593, Abcam), CD42b (ab183345, Abcam) were incubated at 4 °C overnight.

Techniques: Derivative Assay, Immunofluorescence, Expressing, In Vitro, Flow Cytometry, Infection, Enzyme-linked Immunosorbent Assay, Clinical Proteomics