Journal: Cell

Article Title: NLRC5 senses NAD + depletion to form a PANoptosome driving PANoptosis and inflammation

doi: 10.1016/j.cell.2024.05.034

Figure Lengend Snippet: Key Resources Table

Article Snippet: To further improve the AI recognition accuracy, training cycles were repeated until the number of misrecognized regions was reduced to an acceptable level (i.e. until the analysis results did not change appreciably with the completion of further training). . Clinical chemistry analysis Serum creatinine (A11A01933), BUN (A11A01641), iron (A11A01637), ALT (A11A01627), and AST (A11A01629) were detected using ABX Pentra 400 Reagents (HORIBA) and hemoglobin was detected using VetHemaChemRx (Oxford science) according to the manufacturer’s instructions. . Statistical analysis GraphPad Prism 9.0 and 10.0 software were used for in vitro and murine in vivo data analysis.

Techniques: Control, Virus, Recombinant, In Vitro, In Vivo, Protease Inhibitor, Western Blot, Iron Assay, AST Assay, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Isolation, Transfection, Antibody Labeling, Expressing, Software

Journal: Cell

Article Title: NLRC5 senses NAD + depletion to form a PANoptosome driving PANoptosis and inflammation

doi: 10.1016/j.cell.2024.05.034

Figure Lengend Snippet: (A) Survival of 8- to 10-week-old male wild-type (WT) (n = 13) and Nlrc5−/− (n = 10) mice after intraperitoneal injection of phenylhydrazine (PHZ) plus lipopolysaccharide (LPS). (B) Representative images showing hematoxylin and eosin (H&E)-stained kidney sections from WT and Nlrc5−/− mice 30 h after PBS or PHZ plus LPS injection. Outlined areas and arrows denote degenerating kidney tubular epithelium. Images are representative of n = 9 WT (PBS), n = 17 WT (PHZ + LPS), and n = 18 Nlrc5−/− (PHZ + LPS) mice. (C) Serum creatinine levels in WT and Nlrc5−/− mice injected with PBS (WT, n = 9) or PHZ plus LPS (WT, n = 17 and Nlrc5−/−, n = 18) at 30 h post-treatment. (D) Western blot analysis of NLRC5 in kidney tissue from WT mice following treatment with PBS or PHZ plus LPS at 30 h post-treatment. For loading control, α-Tubulin is shown. (E) Densitometric quantification of NLRC5 expression normalized to α-Tubulin expression from WT mice injected with PBS or PHZ plus LPS at 30 h post-treatment (PBS, n = 5; PHZ + LPS, n = 8). (F) Representative images of lipocalin 2 (LCN2) staining in kidney cortex and papilla of WT and Nlrc5−/− mice 30 h after PBS or PHZ plus LPS injection. Images are representative of n = 9 WT (PBS), n = 17 WT (PHZ + LPS), and n = 18 Nlrc5−/− (PHZ + LPS) mice. (G) Western blot analysis of LCN2; pro- and cleaved caspase-3 (CASP3; P35 and P19/P17, respectively); and pro- and cleaved caspase-7 (CASP7; P35 and P20/P17, respectively) in kidney tissue from WT and Nlrc5−/− mice treated with PBS or PHZ plus LPS at 30 h post-treatment. For loading control, β-actin is shown. (H and I) Densitometric quantification of LCN2 (H) and cleaved CASP3 and cleaved CASP7 (I) expression normalized to β-actin expression from WT and Nlrc5−/− mice treated with PBS (WT = 9) or PHZ plus LPS (WT, n = 17 and Nlrc5−/−, n = 18) at 30 h post-treatment. Scale bar = 20 μm (B and F). Mean ± SEM are shown (C, E, H, and I). Nlrc5−/− mice (ref. 56) (A–C and F–I) were used for PHZ plus LPS injection. Two independent experiments were performed to assess survival, and two independent experiments were performed to assess serum parameters, histology, and cell death molecule activation. Pooled results are shown. Statistical analyses were performed using the log-rank (Mantel-Cox) test (A), the one-way ANOVA (C, H, and I), or the unpaired t test (E). *P < 0.05; **P < 0.01; ****P < 0.0001. See also Figure S6.

Article Snippet: To further improve the AI recognition accuracy, training cycles were repeated until the number of misrecognized regions was reduced to an acceptable level (i.e. until the analysis results did not change appreciably with the completion of further training). . Clinical chemistry analysis Serum creatinine (A11A01933), BUN (A11A01641), iron (A11A01637), ALT (A11A01627), and AST (A11A01629) were detected using ABX Pentra 400 Reagents (HORIBA) and hemoglobin was detected using VetHemaChemRx (Oxford science) according to the manufacturer’s instructions. . Statistical analysis GraphPad Prism 9.0 and 10.0 software were used for in vitro and murine in vivo data analysis.

Techniques: Injection, Staining, Western Blot, Control, Expressing, Activation Assay

Journal: Cell

Article Title: NLRC5 senses NAD + depletion to form a PANoptosome driving PANoptosis and inflammation

doi: 10.1016/j.cell.2024.05.034

Figure Lengend Snippet: Key Resources Table

Article Snippet: To further improve the AI recognition accuracy, training cycles were repeated until the number of misrecognized regions was reduced to an acceptable level (i.e. until the analysis results did not change appreciably with the completion of further training). . Clinical chemistry analysis Serum creatinine (A11A01933), BUN (A11A01641), iron (A11A01637), ALT (A11A01627), and AST (A11A01629) were detected using ABX Pentra 400 Reagents (HORIBA) and hemoglobin was detected using VetHemaChemRx (Oxford science) according to the manufacturer’s instructions. . Statistical analysis GraphPad Prism 9.0 and 10.0 software were used for in vitro and murine in vivo data analysis.

Techniques: Control, Virus, Recombinant, In Vitro, In Vivo, Protease Inhibitor, Western Blot, Iron Assay, AST Assay, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Isolation, Transfection, Antibody Labeling, Expressing, Software