Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Differential Interaction of the Staphylococcal Toxins Panton-Valentine Leukocidin and γ-Hemolysin CB with Human C5a Receptors

doi: 10.4049/jimmunol.1500604

Figure Lengend Snippet: (A, B) Amino acid substitutions per 100 amino acids of (A) C5aR1 and (B) C5aR2 of different mammalian species related to human C5aR1 and C5aR2, with a schematic representation of the seven-transmembrane spanning architecture of C5aR1 and C5aR2. (C, D) Amino acid residue alignments of the (C) C5aR1 and (D) C5aR2 of different mammalian species. Extracellular domains are highlighted, and amino acids divergent from the human sequence are shaded. Amino acid sequence alignments were generated by ClustalW alignment using Lasergene MegAlign software (DNAStar). ECL: extracellular loop.

Article Snippet: Amino acid sequence alignments were generated by ClustalW alignment using Lasergene MegAlign software (DNAStar).

Techniques: Sequencing, Generated, Software

Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

Figure Lengend Snippet:

Article Snippet: M(M-CSF), M(LPS/IFN-γ), and M(IL-4) were incubated with human fragment crystallizable receptor (FcR) blocking reagent (Cat # 130059901, Miltenyi Biotec) and extracellularly stained with anti-CD80 BV510 (1:40, clone 2D10, Cat # 305233, Biolegend), anti-CD86 PE-Cy7 (1:40, clone BU63, Cat # 374209, Biolegend), anti-CD206 BC786 (1:125, clone 19.2, Cat # 740999, BD Bioscience), anti-CD209 BV421 (1:66, clone 9E98A, Cat # 330117, Biolegend), anti-HLA-DR APC (1:100, clone L243, Cat # 307622, Biolegend), anti-CD163 PE (1:400, clone GHI/61, Cat # 333606, Biolegend), anti-LRP-1 PE (1:100, clone A2MR-alpha-2, Cat # 12-0919-42, Invitrogen), anti-LRP-2 Alexa Fluor 647 (1:33, clone 545606, Cat # FAB9578R, R&D Systems), anti-EMMPRIN Alexa Fluor 488 (1:800, clone HIM8, Cat # 306207, Biolegend), anti-CD44 APC/Fire 450 (1:40, clone BJ18, Cat # 338818, Biolegend), anti-APP1 Alexa Fluor 488 (1:100, clone 22C11, Cat # MAB348A4, Merck), anti-CD63 APC (1:25, clone H5C6, Cat # 353008, Biolegend), anti-CD74 Superbright 436 (1:40, clone 5–329, Cat # 62-0748-42, Invitrogen), anti-Ku70/80 PE (1:33, clone KU729, Cat # 2547-MSM1-PE-100T, Neobiotechnologies).

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: STAR Protocols

Article Title: In vivo regeneration of neo-nephrons in rodents by renal progenitor cell transplantation

doi: 10.1016/j.xpro.2021.100314

Figure Lengend Snippet:

Article Snippet: Polyclonal goat anti-Megalin antibody , Santa Cruz Biotechnology , Cat# sc-16478; RRID: AB_2234897.

Techniques: Recombinant, Blocking Assay, Gene Expression, TaqMan Assay, Software, Fluorescence, Microscopy

Journal: medRxiv

Article Title: The congenital multiple organ malformation syndrome, Ritscher-Schinzel syndrome is an endosomal recyclinopathy

doi: 10.1101/2024.08.17.24311658

Figure Lengend Snippet: (A) Urinary β2-microglobulin level of individuals with biallelic pathogenic mutations in VPS35L or CCDC93 . (B) The schematic illustration depicts the chimeric constructs of Human-LRP2 utilized in the study. In addition to the wild-type cytoplasmic sequence, mutant constructs were generated in which either or both NPxY motifs were substituted with NPxA. (C) Representative blots of mCherry-nanotrap for mCherry-SNX17 under co-overexpression of chimeric constructs of LRP2. GFP-tagged cytoplasmic tail of CI-MPR, a SNX-BAR cargo protein, was used as negative control. Bar graphs show band intensities relative to WT calculated from three independent experiments. (D) Representative blots for LRP1, LRP2, and N-Cadherin from three independent experiments. Cell surface protein fractions were obtained from HEK293T cell lines. Bar graphs show relative values of KO cells to their rescue or parental cells. (E) Representative view of mCherry-D3 uptake in parental, VPS35L-KO, and VPS35L-rescue cells. Cells were incubated with mCherry-D3 for 30 min, followed by DAPI staining and imaging with a fluorescence microscope. mCherry intensity was quantified using Image J software. 10 fields were acquired in each condition in each of three independent experiments, and mCherry intensity of each field was normalized to the number of DAPI-stained nuclei. Scale bars, 10 µm. (F) HEK293T cell lines were incubated with mCherry-D3 for 30 min followed by FACS analysis to quantitate cellular uptake of mCherry-D3. (G) Immunohistochemistry of Lrp2 in renal tissue in littermate control or Vps35l-cKO Nestin . Red arrows indicate Lrp2 in S1 segment of proximal tubules. Three mice were analyzed in each group. G; Glomerulus. (H) Schematic illustration of the molecular mechanism underlying the proteinuria observed in Ritscher-Schinzel syndrome. (C, D, E) Error bars represent mean± SD. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001.

Article Snippet: The following antibodies were used in this study (WB: western blot, IF: immunofluorescence): rabbit anti-SNX17 (Proteintech, 10275-1-AP, WB), mouse anti-GFP (Roche, 11814460001, WB), rabbit anti-GFP (GeneTex, GTX30738, WB), mouse anti-mCherry (antibodies.com, A85305, WB), rabbit anti-mCherry (antibodies.com, A85306, WB), rabbit anti-CCDC22, (Proteintech, 16636-1-AP, WB), mouse anti-CCDC93, (Origene, CF800568, WB), rabbit anti COMMD4 and rabbit anti COMMD9 (kind gift from Prof. Ezra Burstein, WB) rabbit anti-C16orf62 (Abcam, ab97889, WB), rabbit anti-C16orf62 (Pierce, PA5-28553, IF), rabbit anti-DSCR3 (Merck Millipore, ABN87, WB), rabbit anti-integrin-β1 (Abcam, ab52971, WB), goat anti-VPS35 (antibodies.com, A83699, IF), mouse anti-VPS29 (Santa Cruz, sc-398874, WB), rabbit anti-KIAA1033 (Proteintech, 51101-1-AP, WB), mouse anti-Strumpellin (Santa Cruz, sc-377146, WB), mouse anti-β actin (Sigma, A1978, WB), rabbit anti-LRP1 (Abcam, ab92544, WB), rabbit anti-LRP2 (Proteintech, 19700-1-AP, WB), rabbit anti-LRP2 (prepared as described in , IHC), rabbit anti-LRP4 (SIGMA, HPA012300, WB), rabbit anti-LRP8 (Abcam, ab108208, WB), mouse anti-VLDLR (Santa Cruz, sc-18824, WB), rabbit anti-APP (Abcam, ab32136, WB), rabbit anti-APLP2 (Proteintech, 15041-1-AP, WB), mouse anti-AP (Thermo Fisher, MA1-20245, WB), rabbit anti-Dab1 (kind gift from Dr. M Hattori, IP/WB) , mouse anti-Phosphotyrosine (Merck, 05-321, WB), rabbit anti-GLUT1 (Abcam, ab115730, IF/WB), mouse anti-N-cadherin (Cell signalling technology, 14215S, WB), anti-PSD95 (Merck, MAB1596, WB), mouse anti-FLAG (SIGMA, F1804, WB), rabbit anti-FGFR2, (Proteintech, 13042-1-AP, WB), rabbit anti-ERK1/2 (Cell Signaling Technology, 9102, WB), rabbit anti-phospho-ERK1/2 (Cell Signaling Technology, 9101, WB), anti Ctip2 (Abcam, ab18465, IHC), anti Tbr1 (Abcam, ab275960, IHC), anti Neun (Cell signalling technology, 12943, IHC).

Techniques: Construct, Sequencing, Mutagenesis, Generated, Over Expression, Negative Control, Incubation, Staining, Imaging, Fluorescence, Microscopy, Software, Immunohistochemistry, Control