mouse monoclonal antibody tra 1 85  (R&D Systems)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Degradable fibrin hydrogels for transplantation of iPSC-derived retinal pigment epithelial cell monolayers

doi: 10.3389/fcell.2025.1739620

Figure Lengend Snippet: Histology following RPE debridement and RPE-fibrin transplantation. H&E-stained sections from a control region (A) , debridement zone containing RPE transplant (B) , and debridement zone (C) of Pig 2 sacrificed 2 months after receiving the RPE + fibrin transplant. Transplanted RPE in B exhibit mild depigmentation but are in contact with photoreceptor outer segments (OS) which are shortened relative to control regions. There is also thickening of choroid (Ch), loss of choroidal pigmentation, and cellular infiltration. The debridement zone (C) exhibits significant disruption of retinal layering with tubulation of surviving photoreceptors, choroidal thickening, and absence of RPE. The presence of transplanted RPE was verified by staining for the human specific RPE antibody Tra-1-85 (Green in D–G ) which recognizes CD147. Note Tra-1-85 does not stain RPE in the control region of the pig eye (D) but does stain transplanted cells (E) , RPE in a human eye (F) , and the RPE component of RPE-fibrin in vitro (G) . GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; OS, outer segment; Ch, choroid. Nuclei are stained in (D–G) with DAPI (Blue).

Article Snippet: Immunofluorescence was performed, as described previously ( ; ), using rabbit polyclonal antibody Pab125 to detect Best1 , CD147/EMMPRIN using mouse monoclonal antibody Tra-1-85 (R&D Systems, cat# MAB3195, RRID: AB_2066681), or CRALBP using mouse monoclonal antibody B2 (Novus, cat# NB100-74392, RRID: AB_1048601).

Techniques: Transplantation Assay, Staining, Control, Disruption, In Vitro

Journal: Frontiers in Cell and Developmental Biology

Article Title: SARS-CoV-2 Employ BSG/CD147 and ACE2 Receptors to Directly Infect Human Induced Pluripotent Stem Cell-Derived Kidney Podocytes

doi: 10.3389/fcell.2022.855340

Figure Lengend Snippet: Susceptibility of human iPS cell-derived podocytes to infection by live SARS-CoV-2. qPCR analysis of human iPS cell-derived podocytes infected with SARS-CoV-2 revealed intracellular uptake of the virus for 24 h.p.i (A) , 48 h.p.i (B) and 72 h.p.i (C) . (D) plaque assay quantification from supernatant obtained from infected podocytes at 24, 48 and 72 h.p.i. (E) qPCR analysis of podocyte-specific genes revealed that both synaptopodin (SYNPO) and podocalyxin (PODXL) are significantly upregulated after infection with SARS-CoV-2 at MOI of 0.01, whereas SYNPO is significantly upregulated at multiple MOIs, and PODXL shows no significant changes with viral infection. (F) The expression of spike-associated genes (ACE2, BSG/CD147) and spike processing genes (TMPRSS2, CTSL) are significantly impacted by infection at MOIs of 0.01 and 0.1, respectively. (G) Human iPS cell-derived podocytes treated with SARS-CoV-2 (at MOI of 0.01) immunostain positive for Nucleocapsid protein (magenta), indicating successful infection with the virus. The cells were immunostained also for the podocyte marker Nephrin (green) and counterstained with DAPI (blue). Scale bar: 100 µm (H) Spike positive cells Nephrin and DAPI as nuclear counterstain in the infected podocytes. Scale bar: 100 µm. One-way analysis of variance (ANOVA) with Sidak’s multiple comparison test was used to determine statistical significance. Only p values of 0.05 or lower were considered statistically significant ( p > 0.05 [ns, not significant], p < 0.05 [*], p < 0.01 [**], p < 0.001 [***], p < 0.0001 [****]). Error bars indicate standard deviation of the mean.

Article Snippet: Approximately 1.5 h prior to infecting cells, antibody dilutions were prepared in the CultureBoost-R. We performed the blocking experiment using an ACE2 polyclonal goat antibody (R&D systems; AF933) and CD147 (BSG) mouse monoclonal antibody (Human TRA-1-85/CD147 MAb (Clone TRA-1-85)- R&D systems; MAB3159).

Techniques: Derivative Assay, Infection, Virus, Plaque Assay, Expressing, Marker, Comparison, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: SARS-CoV-2 Employ BSG/CD147 and ACE2 Receptors to Directly Infect Human Induced Pluripotent Stem Cell-Derived Kidney Podocytes

doi: 10.3389/fcell.2022.855340

Figure Lengend Snippet: Transcriptomic and protein level analyses of spike-interacting factors in mature podocytes differentiated from human iPS cells. (A) Heatmap showing expression levels of spike associated genes from three biological human iPS cell-derived podocyte samples. SIGLEC9, Sialic acid-binding Ig-like lectin 9; CAPZA1, F-actin-capping protein subunit alpha-1; CLEC10A, C-type lectin domain family 10 member A; CD33, Myeloid cell surface antigen CD33; TMOD3, Tropomodulin-3; ACE2, Angiotensin Converting Enzyme 2; BSG/CD147, Basigin/CD147 molecule; CD209, CD209 Antigen; MYO6, Unconventional myosin-VI; PLS3, Plastin-3; LDHB, L-lactate dehydrogenase B chain; GNB2L1/RACK, Receptor of activated protein C kinase 1; SNRNP70, U1 small nuclear ribonucleoprotein 70 kDa; DOCK7, Dedicator of cytokinesis protein 7; RPS18, 40S ribosomal protein S18; CAPZB, F-actin-capping protein subunit beta; GOLGA7, Golgin subfamily A member 7; ZDHHC5, Palmitoyltransferase ZDHHC5; SIGLEC10, Sialic acid-binding Ig-like lectin 10; ACTR3, Actin-related protein 3; MYL6, Myosin light polypeptide 6; CORO1C, Coronin-1C; ARPC4, Actin-related protein 2/3 complex subunit 4; CCT6A, T-complex protein 1 subunit zeta; (B) qPCR quantification of nine of the human spike-associated gene from (A) including Transmembrane Serine Protease 2 (TMPRSS2) or cathepsin L (CTSL) in human iPS cell-derived podocytes, Calu-3, Caco-2, glomerular endothelial cells, and HEK 293T cells (normalized to Calu-3 groups). (C) Western blot analysis to evaluate protein expression of ACE2, BSG/CD147, CD209, TMPRSS2 and CTSL in the different cell type used; Calu-3, human iPS cell-derived podocytes, Caco-2, glomerular endothelial (gEndos) cells, and HEK 293T cells. (D) Immunocytochemistry analysis of ACE2, BSG/CD147, CD209, TMPRSS2 and CTSL expression in iPS cell-derived podocytes. Scale bar: 100 µm. One-way analysis of variance (ANOVA) with Sidak’s multiple comparison test was used to determine statistical significance. Only p values of 0.05 or lower were considered statistically significant ( p > 0.05 [ns, not significant], p < 0.05 [*], p < 0.01 [**], p < 0.001 [***], p < 0.0001 [****]). Error bars indicate standard deviation of the mean.

Article Snippet: Approximately 1.5 h prior to infecting cells, antibody dilutions were prepared in the CultureBoost-R. We performed the blocking experiment using an ACE2 polyclonal goat antibody (R&D systems; AF933) and CD147 (BSG) mouse monoclonal antibody (Human TRA-1-85/CD147 MAb (Clone TRA-1-85)- R&D systems; MAB3159).

Techniques: Expressing, Derivative Assay, Binding Assay, Western Blot, Immunocytochemistry, Comparison, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: SARS-CoV-2 Employ BSG/CD147 and ACE2 Receptors to Directly Infect Human Induced Pluripotent Stem Cell-Derived Kidney Podocytes

doi: 10.3389/fcell.2022.855340

Figure Lengend Snippet: Antibody blocking reveal roles of ACE2 and BSG/CD147 receptors in viral uptake. (A) qPCR quantification of S-pseudotyped entry relative to antibody concentration (normalized to unblocked samples). Human iPS cell-derived podocytes were incubated with different dilutions of anti-ACE2, anti-BSG or both for an hour followed by infection with S-pseudotyped virus at MOI 0.02 for 60 h (B) qPCR quantification of Spike binding receptor genes (ACE2, BSG/CD147) and Spike processing factor genes (TMPRSS2, CTSL) at 5 µg/ml for both anti-ACE2 antibody blockage and anti-BSG antibody blockage showing significant changes in gene expression with optimal receptor blockage when compared to unblocked samples (normalized to mock groups). One-way analysis of variance (ANOVA) with Sidak’s multiple comparison test was used to determine statistical significance. Only p values of 0.05 or lower were considered statistically significant ( p > 0.05 [ns, not significant], p < 0.05 [*], p < 0.01 [**], p < 0.001 [***], p < 0.0001 [****]). Error bars indicate standard deviation of the mean.

Article Snippet: Approximately 1.5 h prior to infecting cells, antibody dilutions were prepared in the CultureBoost-R. We performed the blocking experiment using an ACE2 polyclonal goat antibody (R&D systems; AF933) and CD147 (BSG) mouse monoclonal antibody (Human TRA-1-85/CD147 MAb (Clone TRA-1-85)- R&D systems; MAB3159).

Techniques: Blocking Assay, Concentration Assay, Derivative Assay, Incubation, Infection, Virus, Binding Assay, Gene Expression, Comparison, Standard Deviation

Journal: bioRxiv

Article Title: BSG/CD147 and ACE2 receptors facilitate SARS-CoV-2 infection of human iPS cell-derived kidney podocytes

doi: 10.1101/2021.11.16.468893

Figure Lengend Snippet: ( A ) Heatmap showing expression levels of spike associated genes from three biological human iPS cell-derived podocyte samples. SIGLEC9, Sialic acid-binding Ig-like lectin 9; CAPZA1, F-actin-capping protein subunit alpha-1; CLEC10A, C-type lectin domain family 10 member A; CD33, Myeloid cell surface antigen CD33; TMOD3, Tropomodulin-3; ACE2, Angiotensin Converting Enzyme 2; BSG/CD147, Basigin/CD147 molecule; CD209, CD209 Antigen; MYO6, Unconventional myosin-VI; PLS3, Plastin-3; LDHB, L-lactate dehydrogenase B chain; GNB2L1/RACK, Receptor of activated protein C kinase 1; SNRNP70, U1 small nuclear ribonucleoprotein 70 kDa; DOCK7, Dedicator of cytokinesis protein 7; RPS18, 40S ribosomal protein S18; CAPZB, F-actin-capping protein subunit beta; GOLGA7, Golgin subfamily A member 7; ZDHHC5, Palmitoyltransferase ZDHHC5; SIGLEC10, Sialic acid-binding Ig-like lectin 10; ACTR3, Actin-related protein 3; MYL6, Myosin light polypeptide 6; CORO1C, Coronin-1C; ARPC4, Actin-related protein 2/3 complex subunit 4; CCT6A, T-complex protein 1 subunit zeta; ( B ) The expression of Spike associated genes (ACE2, BSG/CD147) and Spike processing genes (TMPRSS2, CTSL) are significantly impacted by infection at MOIs of 0.01 and 0.1, respectively. ( C ) qPCR quantification of nine of the human spike associated gene from (A) including Transmembrane Serine Protease 2 (TMPRSS2) or cathepsin L (CTSL) in human iPS cell-derived podocytes, Calu-3, Caco-2, glomerular endothelial cells, and HEK 293T cells (normalized to Calu-3 groups). ( D ) Western blot analysis to evaluate protein expression of ACE2, BSG/CD147, CD209, TMPRSS2 and CTSL in the different cell type used; Calu-3, human iPS cell-derived podocytes, Caco-2, glomerular endothelial (gEndos)cells, and HEK 293T cells. ( E ) Immunocytochemistry analysis of ACE2, BSG/CD147, CD209, TMPRSS2 and CTSL expression in iPS cell-derived podocytes. Scale bar: 100 µm. One-way analysis of variance (ANOVA) with Sidak’s multiple comparison test was used to determine statistical significance. Only p values of 0.05 or lower were considered statistically significant (p > 0.05 [ns, not significant], p < 0.05 [*], p < 0.01 [**], p < 0.001 [***], p < 0.0001 [****]). Error bars indicate standard deviation of the mean.

Article Snippet: Approximately 1.5 hours prior to infecting cells, antibody dilutions were prepared in the CultureBoost-R. We performed the blocking experiment using an ACE2 polyclonal goat antibody (Cat # AF933; R&D systems) and CD147 (BSG) mouse monoclonal antibody (Human TRA-1-85/CD147 MAb (Clone TRA-1-85)-MAB3159; R&D systems).

Techniques: Expressing, Derivative Assay, Binding Assay, Infection, Western Blot, Immunocytochemistry, Comparison, Standard Deviation

Journal: bioRxiv

Article Title: BSG/CD147 and ACE2 receptors facilitate SARS-CoV-2 infection of human iPS cell-derived kidney podocytes

doi: 10.1101/2021.11.16.468893

Figure Lengend Snippet: ( A ) qPCR quantification of S-pseudotyped entry relative to antibody concentration (normalized to unblocked samples). Human iPS cell-derived podocytes were incubated with different dilutions of anti-ACE2, anti-BSG or both for an hour followed by infection with S-pseudotyped virus at MOI 0.02 for 60 h. ( B ) qPCR quantification of Spike binding receptor genes (ACE2, BSG/CD147) and Spike processing factor genes (TMPRSS2, CTSL) at 5 µg/ml for both anti-ACE2 antibody blockage and anti-BSG antibody blockage showing significant changes in gene expression with optimal receptor blockage when compared to unblocked samples (normalized to mock groups). One-way analysis of variance (ANOVA) with Sidak’s multiple comparison test was used to determine statistical significance. Only p values of 0.05 or lower were considered statistically significant (p > 0.05 [ns, not significant], p < 0.05 [*], p < 0.01 [**], p < 0.001 [***], p < 0.0001 [****]). Error bars indicate standard deviation of the mean.

Article Snippet: Approximately 1.5 hours prior to infecting cells, antibody dilutions were prepared in the CultureBoost-R. We performed the blocking experiment using an ACE2 polyclonal goat antibody (Cat # AF933; R&D systems) and CD147 (BSG) mouse monoclonal antibody (Human TRA-1-85/CD147 MAb (Clone TRA-1-85)-MAB3159; R&D systems).

Techniques: Concentration Assay, Derivative Assay, Incubation, Infection, Virus, Binding Assay, Gene Expression, Comparison, Standard Deviation