Journal: Animals : an Open Access Journal from MDPI

Article Title: Mechanobiological Strategies to Enhance Ovine ( Ovis aries ) Adipose-Derived Stem Cells Tendon Plasticity for Regenerative Medicine and Tissue Engineering Applications

doi: 10.3390/ani14152233

Figure Lengend Snippet: Summary of antibodies used in cytofluorimetric analysis.

Article Snippet: Class-I HLA , FITC , Novus Biologicals , Cambridge, UK.

Techniques:

Journal: eLife

Article Title: Multimodal HLA-I genotype regulation by human cytomegalovirus US10 and resulting surface patterning

doi: 10.7554/eLife.85560

Figure Lengend Snippet: ( A ) Wild-type HeLa cells or TagBFP-T2A-US10i clone #5 were treated with doxycycline (0.33 µg/mL) or DMSO for 24 hr. Subsequently. cells were stained by anti-CD71, anti-Bw6, or anti-HLA-C antibodies and a flow cytometry analysis was performed (cells were gated according to TagBFP expression, TagBFP + or TagBFP - ). ( B ) Experimental scheme of pSILAC immunopeptidomics. HeLa TagBFP-T2A-US10i #5 was pulse-labeled with medium-heavy (M) or heavy (H) amino acids in the presence of DMSO or doxycycline, respectively. After 24 hr, metabolically labeled cells were harvested and combined in a 1:1 ratio. HLA-I peptides were isolated and analyzed by nanoLC-MS/MS. The scheme was created with BioRender.com ( C ) Scatter plots showing median log2 H/M ratios and log10 intensity of quantified HLA-I peptides with respect to their HLA-I allele. Dot size correlates with number of ratios used to calculate the corresponding peptide ratio. ( D ) HLA-I consensus binding motif of HLA-B*15:03 obtained from NetMHCpan 4.1 motif viewer ( https://services.healthtech.dtu.dk/services/NetMHCpan-4.1/ ) ( E ) Gibbs clustering analysis (GibbsCluster 2.0) for quantified HLA-B*15:03 peptides with median log2 H/M ratios ≤0 or >0.

Article Snippet: The following antibodies were used in this study: W6/32 (anti-pan-HLA-I assembled with β 2 m and peptide; ), anti-HLA-B*07:02 (BB7.1; ), anti-HLA-A*02 (BB7.2; ), anti-β 2 m (BBM.1; ), anti-HLA-B*44 (TT4-A20; ), anti-HLA-A*03 (GAP A3; ), anti-HLA-C (DT-9, BioLegend, 373302, RRID :AB_2650941 ), anti-HLA-E (3D12, BioLegend, 342602, RRID :AB_1659247 ), anti-HA produced in mouse (Sigma-Aldrich, H3663, RRID: AB_262051 ) or rabbit (Sigma-Aldrich, H6908, RRID :AB_260070 ), anti-ERp57 (abcam, ab13506, RRID :AB_1140700 ), anti-β-actin (Sigma-Aldrich, A2228, RRID :AB_476697 ), anti-CD28 (BD Biosciences), APC-coupled anti-HA (Miltenyi Biotec, 130-098-404, RRID :AB_2751024 ), APC-coupled IgG1-isotype control (Miltenyi Biotec, 130-113-200, RRID :AB_2733881 ), APC-coupled anti-CD8 (BD Biosciences), APC-coupled anti-CD56 (Miltenyi Biotec, 130-113-305, RRID :AB_2726084 ), APC-coupled goat anti-mouse IgG (BD Biosciences, 550826, RRID :AB_398465 ), FITC-coupled anti-IFNγ (BD Biosciences, 554551, RRID: AB_395473 ), PE/Cy7-coupled anti-TNFα (BioLegend, 506323, RRID :AB_2204356 ), HRP-coupled goat anti-mouse IgG (Dianova, 115-035-146, RRID :AB_2307392 ), HRP-coupled goat anti-rabbit IgG (Sigma-Aldrich, 12-348, RRID :AB_390191 ), FITC-coupled anti-Bw6 (Miltenyi Biotec, 130-123-264, RRID: AB_2819460 ), and APC-coupled anti-CD71 (Miltenyi Biotec, 130-123-788, RRID :AB_2857636 ).

Techniques: Staining, Flow Cytometry, Expressing, Labeling, Metabolic Labelling, Isolation, Tandem Mass Spectroscopy, Binding Assay

Journal: Cellular and Molecular Life Sciences

Article Title: iRhom2 regulates ectodomain shedding and surface expression of the major histocompatibility complex (MHC) class I

doi: 10.1007/s00018-024-05201-7

Figure Lengend Snippet: List of the significantly ( p value < 0.05) reduced potentially shed proteins detected in the secretome of LPS/PMA stimulated iRhom2 KO BMDMs versus WT controls

Article Snippet: Briefly, HTB94 were stained with FITC anti-HLA Class I [W6/32], isotype IgG2a (Abcam, Cambridge, UK) or PE anti-HLA Class I [W6/32], isotype IgG2a (BioLegend, San Diego, CA) at room temperature for 15 min according to the manufacturer’s instruction, washed with PBS and then resuspended in 300 µl PBS.

Techniques:

Journal: Cellular and Molecular Life Sciences

Article Title: iRhom2 regulates ectodomain shedding and surface expression of the major histocompatibility complex (MHC) class I

doi: 10.1007/s00018-024-05201-7

Figure Lengend Snippet: List of the significantly ( p value < 0.05) reduced proteins detected in the secretome of iRhom2 KO BMDMs versus WT controls in steady-state cnditions

Article Snippet: Briefly, HTB94 were stained with FITC anti-HLA Class I [W6/32], isotype IgG2a (Abcam, Cambridge, UK) or PE anti-HLA Class I [W6/32], isotype IgG2a (BioLegend, San Diego, CA) at room temperature for 15 min according to the manufacturer’s instruction, washed with PBS and then resuspended in 300 µl PBS.

Techniques:

Journal: Cellular and Molecular Life Sciences

Article Title: iRhom2 regulates ectodomain shedding and surface expression of the major histocompatibility complex (MHC) class I

doi: 10.1007/s00018-024-05201-7

Figure Lengend Snippet: List of the significantly ( p value < 0.05) increased potentially shed proteins detected in the secretome of stimulated PBMC-derived macrophages versus untreated controls

Article Snippet: Briefly, HTB94 were stained with FITC anti-HLA Class I [W6/32], isotype IgG2a (Abcam, Cambridge, UK) or PE anti-HLA Class I [W6/32], isotype IgG2a (BioLegend, San Diego, CA) at room temperature for 15 min according to the manufacturer’s instruction, washed with PBS and then resuspended in 300 µl PBS.

Techniques:

Journal: Cellular and Molecular Life Sciences

Article Title: iRhom2 regulates ectodomain shedding and surface expression of the major histocompatibility complex (MHC) class I

doi: 10.1007/s00018-024-05201-7

Figure Lengend Snippet: List of the significantly ( p value < 0.05) increased potentially shed proteins detected in the secretome of LPS/PMA stimulated BMDMs versus untreated controls

Article Snippet: Briefly, HTB94 were stained with FITC anti-HLA Class I [W6/32], isotype IgG2a (Abcam, Cambridge, UK) or PE anti-HLA Class I [W6/32], isotype IgG2a (BioLegend, San Diego, CA) at room temperature for 15 min according to the manufacturer’s instruction, washed with PBS and then resuspended in 300 µl PBS.

Techniques: