Journal: Scientific reports

Article Title: Complement classical and alternative pathway activation contributes to diabetic kidney disease progression: a glomerular proteomics on kidney biopsies.

doi: 10.1038/s41598-024-84900-4

Figure Lengend Snippet: Fig. 3. Immunohistochemistry (IHC) staining for C1q, C3, C4, C5b-9, CFB, CFH, MBL, and MASP. (a–e) Representative images of C1q, C3, C4, C5b-9, and CFB staining showing a GBM pattern of immunostaining in kidney tissues from patients with DKD. (f) Immunohistochemical diagram of CFH in kidneys with DKD depicting mesangial deposition. (g, h) Representative images of MBL and MASP staining showing no deposition was found in the glomeruli of kidneys with DKD. Scale bars represent 50 μm. CFB, complement factor B; CFH, complement factor H; MBL, mannose-binding lectin; MASP, mannose-binding lectin- associated serine protease; GBM, glomerular basement membrane; DKD, diabetic kidney disease.

Article Snippet: After washing, sections were incubated with either anti-C1q antibodies (1:200 dilution, 16889-1-AP; Proteintech), anti-C3 antibodies (1:200 dilution, ab200999; Abcam), anti-C4 antibodies (1:300 dilution, 22233-1-AP; Proteintech), anti-C5b-9 antibodies (1:400 dilution, ab55811; Abcam), anti-MBL antibodies (1:200 dilution, ab23457; Abcam), anti-MASP1 antibodies (1:400 dilution, 21837-1-AP; Proteintech), anti-CFB antibodies (1:50 dilution, ab192577; Abcam), or anti-CFH antibodies (1:50 dilution, ab170036; Abcam) overnight at 4 °C in a moist chamber.

Techniques: Immunohistochemistry, Staining, Immunostaining, Immunohistochemical staining, Binding Assay, Membrane

Journal: Nature

Article Title: LACC1 bridges NOS2 and polyamine metabolism in inflammatory macrophages

doi: 10.1038/s41586-022-05111-3

Figure Lengend Snippet: NOS2 is indispensable for the antibacterial function of LACC1. a, Illustration of the NOS2-LACC1-ODC1 signaling axis. b, Body weight of each mouse was monitored daily after S . Typhimurium infection (n = 10, P = 0.0414). c, Mouse survival curves (n = 10). d, CFUs in fecal pellets were measured at day 4 after infection. e, f, g, CFUs in caecum, spleen and liver, respectively, were measured at day 6 after infection. h, i, j, ELISA measurements of TNFα, IL6 and IL12b, respectively, secreted by BMDMs stimulated by LPS and IFNγ with or without 1 mM DFMO treatment. k, CFUs of intracellular S . Typhimurium in inflammatory BMDMs. l, LDH assay in S . Typhimurium infected inflammatory BMDMs. The mean and SEM (error bars) are derived from three ( - ) or ten ( - ) biological replicates (n = 3 or n = 10). Statistical significance (two-tailed t-test) compared to control (Ctrl): *P<0.05; **P<0.01; ns, not significant.

Article Snippet: In some experiments as indicated, 1 mM DFMO (Cayman, 16889) was supplemented during differentiation to inhibit ODC1.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Derivative Assay, Two Tailed Test, Control

Journal: Nature

Article Title: LACC1 bridges NOS2 and polyamine metabolism in inflammatory macrophages

doi: 10.1038/s41586-022-05111-3

Figure Lengend Snippet: Irreversible inhibition of ODC1 phenocopied exacerbated S . Typhimurium induced cell death from Lacc1 −/− inflammatory BMDMs. a, b, c, ELISA measurements of TNFα, IL6, and IL12b, respectively, secreted by BMDMs stimulated by LPS and IFNγ with or without 1 mM DFMO and/or 500 μM putrescine (Put) treatment. d, CFUs of intracellular S . Typhimurium in inflammatory BMDMs. e, LDH assay in S . Typhimurium infected inflammatory BMDMs. The mean and SEM (error bars) are derived from three biological replicates (n = 3). Statistical significance (two-tailed t-test) compared to control (Ctrl): *P<0.05; **P<0.01; ns, not significant.

Article Snippet: In some experiments as indicated, 1 mM DFMO (Cayman, 16889) was supplemented during differentiation to inhibit ODC1.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Infection, Derivative Assay, Two Tailed Test, Control