recombinant clu protein  (MedChemExpress)

Journal: Nature Communications

Article Title: Podocyte-specific KLF6 primes proximal tubule CaMK1D signaling to attenuate diabetic kidney disease

doi: 10.1038/s41467-024-52306-5

Figure Lengend Snippet: a , b OCR and quantifications in 1° PT treated with conditioned media (CM) from 1° KLF6 PODTA (OE) versus NPHS2-rtTA (WT) podocytes in normal glucose (NG) and high glucose (HG). n (biological replicates) = 7; basal respiration: p = 0.0013:OE CM (NG) vs WT CM (HG), p = 0.0008:OE CM (NG vs HG), p = 0.0002:WT CM vs OE CM (HG); ATP production: p = 0.0006:OE CM (NG) vs WT CM (HG), p = 0.0004:WT CM (NG vs HG), p = < 0.0001:WT CM vs OE CM (HG); maximal respiration: p = 0.0343:WT CM (NG vs HG), p = 0.0389:OE CM (NG) vs WT CM (HG); spare respiratory capacity: p = 0.0143:OE CM (NG) vs WT CM (HG), p = 0.0117:WT CM (NG vs HG). c , d Volcano plot showing podocyte secretome and urine proteome from mice ( n = 3). e Circos plot highlighting ligand-receptor interactions for upregulated proteins in podocyte secretome and DEGs in preconditioned-PT from snRNA-seq. f Representative images of ApoJ immunohistochemistry. Arrows-ApoJ staining ( n = 3 mice/group, scale bar = 25 µm). g , h OCR and quantifications in 1° PT cells treated with CM + anti-ApoJ blocking antibody or CM + anti-IgG in HG ( n (biological replicates) = 7); WT CM vs OE CM: p = 0.0259(basal respiration), p = 0.0026(maximal respiration), p = 0.0004(spare respiratory capacity). i , j OCR and quantifications in 1° PT treated with CM from differentiated human podocytes in HG ( n (biological replicates) = 6, p = 0.0001(basal respiration), p = 0.0009(ATP production), p = 0.005(maximal respiration), p = 0.0156(spare respiratory capacity)). k Western blot for immunoprecipitation of ApoJ in 1° PT treated with CM in NG/HG. l , m Representative images from co-immunostaining for ApoJ, CaMK1D, Lrp2, and Hoechst. Arrows-indicate ApoJ/CamK1D and ApoJ/Lrp2 colocalization. (scale bar = 50 µm, 5 µm for inset). n , o OCR and quantification in 1° PT treated with recombinant ApoJ ± cilastatin/VEH ( n (biological replicates) = 7 for control), ( n (biological replicates)=5) for ApoJ+VEH, (n(biological replicates) = 5) for ± cilastatin; basal respiration: p = 0.0003(control vs ApoJ + VEH), p = 0.0157(control vs cilastatin), p = 0.0329 (ApoJ + VEH vs ApoJ + cilastatin); ATP production: p = 0.0001 (control vs ApoJ + VEH), p = 0.0086 (ApoJ + VEH vs ApoJ + cilastatin); maximal respiration: p = 0.0151 (control vs ApoJ + VEH). p Proposed schematic of potential KLF6-ApoJ-CaMK1D signaling between podocytes and PT cells. For all data: * p < 0.05, ** p < 0.01 *** p < 0.001, Kruskal–Wallis test with Dunn’s posttest; data presented as mean ± SEM. Source data are provided as a Source Data file. p Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en .

Article Snippet: Mouse 1° PT cells were treated with recombinant mouse ApoJ-His-tag (His-ApoJ) (20 µg/ml) (SinoBiological, 50485-M08H) and Cilastatin (0.1 mg/ml) (MedChemExpress, HY-A0166A), for 24 h under NG conditions.

Techniques: Immunohistochemistry, Staining, Blocking Assay, Western Blot, Immunoprecipitation, Immunostaining, Recombinant, Control

Journal: Acta Neuropathologica Communications

Article Title: Clusterin ameliorates tau pathology in vivo by inhibiting fibril formation

doi: 10.1186/s40478-020-01079-1

Figure Lengend Snippet: CLU binds to tau and prevents tau aggregation. a In vitro tau fibrilization assay was performed with the 4R0N tau isoform in the presence or absence of recombinant human CLU and thioflavine T intensity was monitored during 24-h time course. b Quantification of thioflavine T signal at the 24-h time point. Four replicates were used. Data present as mean ± S.E.M. and analyzed by Student’s t test *** p < 0.001. c Western blot analysis of soluble (supernatant, s) and insoluble (pellet, p) fractions following tau fibril assembly in the presence or absence of exogenous CLU. d Tau assembly assay performed with 4R0N recombinant tau, tau PHFs from AD brains, and increasing concentrations of recombinant CLU. Four replicates were used. Data present as mean ± S.E.M. and analyzed by one-way ANOVA, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Tau filament assembly was performed with 5 μM 4R0N isoform in assembly buffer (10 mMHEPES (pH7.4), 33 mM NaCl, 1 mM MgCl2, 1.5 mM EGTA, 60 μM EDTA, 5uM Thioflavin-T and 20ug/ml Heparin) with and without 4 μM recombinant CLU (R&D #2937-HS-050).

Techniques: In Vitro, Recombinant, Western Blot