primary antibody  (ProSci Incorporated)

Journal: Science Advances

Article Title: Discovery of SARS-CoV-2 papain-like protease (PL pro ) inhibitors with efficacy in a murine infection model

doi: 10.1126/sciadv.ado4288

Figure Lengend Snippet: In each panel, SARS-CoV-2 PL pro bound to 4 (sticks, salmon) is shown in cartoon representation, color-coded by subdomain, and with light gray surface representation. ( A ) Structural superimposition of SARS-CoV-2 PL pro bound to 4 and SARS-CoV-2 PL pro bound to mouse ISG15 [yellow; Protein Data Bank (PDB) ID 6YVA]. The PL pro protein structure in 6YVA is not shown for clarity. The C terminus of ISG15 (His 149 -Gly 155 ) overlaps with the binding site of 4 . The catalytic triad is shown in magenta; a catalytically dead construct of PL pro (C111S) was used to generate the crystal structure. ( B ) Detailed view of 4 binding in the BL2 loop of SARS-CoV-2 PL pro , highlighting critical protein-ligand interactions. Hydrogen bonds are represented as black dashed lines. Tyr 264 is not labeled but is in the background, forming a hydrogen bond with the amide of 4 ( C to E ) Structural overlays of SARS-CoV-2 PL pro bound 4 and SARS-CoV-2 bound to GRL0617 (wheat; PDB ID 7CMD). The orientation of the binding pocket varies in each panel, with the BL2 loop labeled throughout. Surface representation is shown for the structure of SARS-CoV-2 PL pro bound to 4 only.

Article Snippet: Next, the tissue sections were incubated for 30 min with SARS-CoV-2 (COVID-19) nucleocapsid antibody (HL448; GTX635686, GeneTex) at a 1:10,000 dilution, followed by incubation for 30 min with DAB (3,3’-diaminobenzidine) rabbit secondary reagents (Bond Polymer Refine Detection, DS9800, Leica Biosystems).

Techniques: Binding Assay, Construct, Labeling

Journal: Science Advances

Article Title: Discovery of SARS-CoV-2 papain-like protease (PL pro ) inhibitors with efficacy in a murine infection model

doi: 10.1126/sciadv.ado4288

Figure Lengend Snippet: ( A ) Dose-response curve of 4 assayed in a human airway epithelial cell model of SARS-CoV-2 infection . The fraction unbound of 4 in the dNHBE medium was measured to be 0.916, assumed to be 1 for modeling purposes. ( B ) Profiling 4 through safety panels and CYP panels reveal no clear off-target liabilities. See table S1 for full Cerep profile. ( C ) In vitro and in vivo absorption, distribution, and metabolism properties across multiple species. MDR, multi-drug resistance protein 1 (P-glycoprotein) efflux ratio; BCRP, murine Breast Cancer Resistance Protein BA/AB ratio; PPB, Plasma Protein Binding.

Article Snippet: Next, the tissue sections were incubated for 30 min with SARS-CoV-2 (COVID-19) nucleocapsid antibody (HL448; GTX635686, GeneTex) at a 1:10,000 dilution, followed by incubation for 30 min with DAB (3,3’-diaminobenzidine) rabbit secondary reagents (Bond Polymer Refine Detection, DS9800, Leica Biosystems).

Techniques: Infection, In Vitro, In Vivo, Protein Binding

Journal: Science Advances

Article Title: Discovery of SARS-CoV-2 papain-like protease (PL pro ) inhibitors with efficacy in a murine infection model

doi: 10.1126/sciadv.ado4288

Figure Lengend Snippet: ( A ) Dose-range finding studies showed that the selected doses cover a range of trough concentrations from subtherapeutic to therapeutic levels. ( B ) Representative immunohistochemistry images from lung histopathology of mouse-adapted SARS-CoV-2 infection 4 days after dose of 4 at 20, 50, and 150 mg/kg; Nirmatrelvir at 1000 mg/kg; vehicle; or uninfected, untreated animals. ( C ) Compound 4 protected mice from weight loss, with roughly 10% weight loss seen in the vehicle arm consistent with data in . PO, by mouth/oral administration; *** P < 0.001; **** P < 0.0001; n.s., not significant. ( D ) Compound 4 led to a statistically significant and dose-dependent reduction in day 4 lung viral titers. Titers were plotted as mean log 10 50% cell culture infectious dose (CCID 50 )/ml ± SEM, with data analysis and significance levels matching those in .

Article Snippet: Next, the tissue sections were incubated for 30 min with SARS-CoV-2 (COVID-19) nucleocapsid antibody (HL448; GTX635686, GeneTex) at a 1:10,000 dilution, followed by incubation for 30 min with DAB (3,3’-diaminobenzidine) rabbit secondary reagents (Bond Polymer Refine Detection, DS9800, Leica Biosystems).

Techniques: Immunohistochemistry, Histopathology, Infection, Cell Culture

Journal: Science Advances

Article Title: Discovery of SARS-CoV-2 papain-like protease (PL pro ) inhibitors with efficacy in a murine infection model

doi: 10.1126/sciadv.ado4288

Figure Lengend Snippet: ( A ) The location of the cysteine protease, PL pro in SARS-CoV-2 genome, and the cleavage sites that it processes to generate functional viral proteins. ( B ) The Nsp3 protein (residues 1 to 1944) showing all domains identifying clinically observed Omicron mutations with frequency > 1%. ( C ) PL pro domain (residues using PL pro numbering 3 to 315) with location of mutation with frequency > 0.5% in green, which are distal from residues within 5.5 Å for 4 (pink spheres). Binding surface is shown in as blue and cyan, where cyan indicates the three highest frequency mutations (A246, P347, and K157) within the ligand binding site (table S8). ( D ) Structural superposition of ISG15 binding site and 4 showing interaction with similar residues.

Article Snippet: Next, the tissue sections were incubated for 30 min with SARS-CoV-2 (COVID-19) nucleocapsid antibody (HL448; GTX635686, GeneTex) at a 1:10,000 dilution, followed by incubation for 30 min with DAB (3,3’-diaminobenzidine) rabbit secondary reagents (Bond Polymer Refine Detection, DS9800, Leica Biosystems).

Techniques: Functional Assay, Mutagenesis, Binding Assay, Ligand Binding Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: SARS-CoV-2 directly infects the pancreatic islets of non-human primates (NHPs). a Representative multi-label IF image from the elder control NHP sample was stained for SARS-CoV-2 S1 protein (S, green), glucagon (α, red), insulin (β, cyan), somatostatin (δ, magenta), and polypeptide (P, yellow). Scale bars, 50 μm. b – d Pancreatic tissue section from one SARS-CoV-2-infected elder NHP sample was stained by the same panel of multi-label IF, showing the co-localization of S and P. Scale bars, 800 μm. c , d Representative multi-label IF image from the magnified section of ( b ). Inset highlights SARS-CoV-2 viral antigen co-localized with islet endocrine cells. Scale bars, 20 μm. e Pancreatic tissue section from another SARS-CoV-2-infected elder NHP sample was stained by the same panel of multi-label IF, showing the co-localization of S and markers of various islet endocrine cells. Scale bars, 800 μm. f Representative multi-label IF image from the magnified section of ( e ). Inset highlights co-expression of SARS-CoV-2 S protein (S, green) with glucagon (α, red), insulin (β, cyan), somatostatin (δ, magenta), and polypeptide (P, yellow). Scale bars, 100 μm. g – j Quantification of the percentage of glucagon + α, insulin + β, somatostatin + δ, and polypeptide + PP cells, as well as SARS-CoV-2 S protein + glucagon + α, S protein + insulin + β, and S protein + somatostatin + δ cells, in the elder control NHPs (3 slides) and elder COVID-19 model NHPs(4 slides) ( n = 10 images examined from all slides/group). Data are presented as mean ± SD. p Values were calculated by paired or unpaired two-tailed Student’s t test. *p < 0.05, **p < 0.01, and ***p < 0.001

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Staining, Infection, Expressing, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: SARS-CoV-2 directly infects the human pancreatic islet cells. a Pancreatic tissue sections from the deceased control subject were stained by multi-label IF for SARS-CoV-2 S1 protein (S, green), glucagon (α, red), insulin (β, cyan), and somatostatin (δ, yellow) ( n = 10 images examined in total). Scale bars, 50 μm. b Pancreatic tissue sections from the COVID-19 autopsy samples (the prototypic SARS-CoV-2 strain-infected) were stained by multi-label IF for SARS-CoV-2 S1 protein (S, green), glucagon (α, red), insulin (β, cyan), and somatostatin (δ, yellow) ( n = 10 images examined in total). Scale bars, 200 μm. c Representative multi-label IF image from the magnified section of ( b ). Inset highlights SARS-CoV-2 viral antigen scattered in a severely damaged and necrotic islet. Scale bars, 50 μm. c The serial section from the same pancreatic tissue stained for H&E and the same necrotic islet was magnified and circled (blue) to clearly observe its pathological changes. The islet structure is disorganized, with cellular swelling and degeneration observed in the internal regions. Cytoplasmic eosinophilia is intensified, and nuclear swelling is evident. The pentagon-marked area (☆) indicates significant necrosis of islet cells, with nuclear pyknosis, karyolysis, and loss of original cellular structure, leaving only homogeneously stained eosinophilic areas. d , e Representative multi-label IF image from the magnified section of (a). Inset highlights SARS-CoV-2 viral antigen co-localized with islet endocrine cells. Scale bars, 50 μm. COVID-19, coronavirus disease 2019; H&E, hematoxylin and eosin; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2. f – i Quantification of the percentage of glucagon + α, insulin + β, somatostatin + δ, and SARS-CoV-2 S protein + cells, as well as S protein + glucagon + α, S protein + insulin + β, and S protein + somatostatin + δ cells, in control human pancreatic tissues and the COVID-19 patients pancreatic tissues ( n = 10 images/group). Data are presented as mean ± SD. p Values were calculated by paired or unpaired two-tailed Student’s t test. *p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Staining, Infection, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: Characteristics of pathological changes in adult and elder COVID-19 NHPs and vaccination triggers insulin receptor activation in the COVID-19 NHP model. a The main lesions in the pancreatic islets observed in the elder prototypic SARS-CoV-2 strain-infected model (a-3 to a-5) compared with the adult (a-1) and elder control (a-2) animals. The first line shows the representative images of pancreatic tissue sections stained by H&E. The second line shows the representative IHC images of pancreatic tissue sections stained for insulin (red) and glucagon (brown). (a-1) Normal pancreatic islets and surrounding tissues in the adult control group; (a-2) a few islet cells are swelling of individual pancreatic islets in the elder control group. In the elder prototypic SARS-CoV-2 strain-infected model, (a-3) the number of islets cells is significantly decreased, and a large amount of amyloid substance is deposited in some islets; (a-4) glucagon + alpha cells are increased in individual islets; (a-5) a few islets are atrophy. Scale bars, 50 μm or 100 μm shown in the corresponding images below. b The main lesions in the exocrine pancreas observed in the elder prototypic SARS-CoV-2 strain-infected model (b-1 to b-5). (b-1) Exfoliated cells can be observed in a few pancreatic duct lumen; (b-2) hyperplasia of local pancreatic ductal; (b-3) dilation of pancreatic duct; (b-4) pancreatic periductal inflammatory cells infiltrated around the pancreatic duct; (b-5) inflammatory cells infiltrated in the interstitium. The representative images of pancreatic tissue sections stained by H&E. Scale bars, 50 μm or 100 μm shown in the corresponding images below. c Qualitative and quantitative analysis by Congo red staining to examine prototypic SARS-CoV-2 strain-infection-elicited islet amyloidosis in the elder COVID-19 model (4 slides) compared with the elder control (3 slides) ( n = 10 images examined from all slides/group). d The serial section was stained by Masson’s trichrome staining, qualitatively and quantitatively, to observe SARS-CoV-2-infection-elicited islet amyloidosis in the elder COVID-19 model infected with the prototypic SARS-CoV-2 strain compared with the elder control ( n = 10 images examined in total/group). e – h Representative multi-label IF image from adult control ( a ), prototypic SARS-CoV-2-strain-infected COVID-19 NHP model ( b ), and vaccinated COVID-19 NHP model ( c , d ) pancreatic tissue section samples were stained for ICA512 (I5, red), insulin receptor α (IRα, yellow), insulin receptor β (IRβ, magenta), CK19 (CK, white), and trypsin (T, cyan). Scale bars, 20 μm in ( a – c ). c Representative multi-label IF images from section ( d ). Scale bars, 100 μm in ( d ). i Quantification of the percentage of ICA512 + , insulin receptor α (Ins R α) + , insulin receptor β (Ins R β) + , CK19 + , and trypsin + cells in the control NHPs (3 slides), COVID-19 NHP model (3 slides) and vaccinated COVID-19 NHP model (3 slides) ( n = 10 images examined from all slides /group). Data are presented as mean ± SD. p Values were calculated by paired or unpaired two-tailed Student’s t test. *p < 0.05, **p < 0.01, and ***p < 0.001

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Activation Assay, Infection, Staining, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: Characteristics of the markers of microvascular damage and cellular stress in elder control and elder COVID-19 model NHPs. a , b The markers of microvascular damage and cellular stress, ICAM-1 and G3BP1, were elevated in the islets of elder COVID-19 model ( a ) compared with the elder control NHPs ( b ). Representative multi-labels IF image in the pancreas from the elder control samples were stained for glucagon (α, magenta), insulin (β, green), ICAM-1 (IC, yellow), G3BP1 (G3, cyan), VCAM-1 (VC, red) and Cleaved caspase 3 (Cas3, white). Scale bars, 20 μm. c – g Quantification of the percentage of ICAM-1 + cell, G3BP1 + cell, VCAM-1 + cell, and cleaved caspase 3 + cell, as well as glucagon + insulin + cell, ICAM-1 + insulin + cell, G3BP1 + insulin + cell and VCAM-1 + insulin + cell in the elder control NHPs (3 slides) and elder COVID-19 model NHPs (4 slides), ( n =10 images examined from all sildes/group). Data are presented as mean ± SD. p values were calculated by unpaired two-tailed Student’s t test. * p < 0.05 and *** p < 0.001

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Staining, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: SARS-CoV-2 infection aggregated activation of α-SMA and accumulation of collagen fibers from the NHP pancreatic tissues of the elder COVID-19 model. Pancreatic tissue sections from the elder control NHP samples were stained by multi-label immunofluorescence (IF) for COL1A1 (CL, yellow), α-SMA (α, red), Ki67 (K, magenta), SARS-CoV-2 S1 protein (S, cyan), insulin (β, green), and CD31 (CD, white). Scale bars, 200 μm. Representative multi-label IF image from the magnified section of ( a ). Scale bars, 20 μm. The pancreatic tissue section from one elder NHP model sample was stained by the same multi-label makers in ( a ). Scale bars, 400 μm. Representative multi-label IF image from the magnified section of ( c ). Inset highlights proliferating collagen fibers dividing the exocrine and endocrine pancreatic tissues into various islands. Scale bars, 20 μm. e Pancreatic tissue section from another elder prototypic SARS-CoV-2 strain-infected NHP model sample was stained by the same multi-label makers in ( a ). Scale bars, 800 μm. f , g Representative multi-label IF image from the magnified section of ( e ). Inset highlights co-localization of SARS-CoV-2 viral antigen and accumulated collagen fibers in damaged islet insulin + β cells. Scale bars in f 40 μm. Scale bars in g 20 μm. h – k Quantification of the percentage of COL1A1 + , α-SMA + , Ki67 + , and CD31 + cells, as well as insulin + COL1A1 + , insulin + α-SMA + , and S protein + CD31 + cells, in the elder control NHPs (3 slides) and elder COVID-19 NHP model (4 slides) ( n = 10 images examined from all slides /group). Data are presented as mean ± SD. p Values were calculated by unpaired two-tailed Student’s t test. *p < 0.05, **p < 0.01, and ***p < 0.001

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Infection, Activation Assay, Staining, Immunofluorescence, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: SARS-CoV-2 infection aggregated activation of α-SMA and accumulation of collagen fibers in the human pancreas. a Pancreatic tissue sections from deceased control subject were stained by multi-label immunofluorescence (IF) for COL1A1 (CL, yellow), α-SMA (α, red), Ki67 (K, magenta), SARS-CoV-2 S1 protein (S, cyan), insulin (β, green), and CD31 (CD, white). Scale bars, 800 μm. b Pancreatic tissue sections from the prototypic SARS-CoV-2 strain-infected COVID-19 autopsy samples were stained by multi-label immunofluorescence (IF) for COL1A1 (CL, yellow), α-SMA (α, red), Ki67 (K, magenta), SARS-CoV-2 S1 protein (S, cyan), insulin (β, green), and CD31 (CD, white). Scale bars, 800 μm. c – e Representative multi-label IF image from the magnified section of ( a ). Inset highlights activation of α-SMA and proliferation of collagen fibers in both the exocrine and endocrine pancreatic tissues. Scale bars, 50 μm. f – i Quantification of the percentage of COL1A1 + , α-SMA + , Ki67 + , and CD31 + cells, as well as insulin + COL1A1 + , insulin + α-SMA + , and S protein + CD31 + cells, in the control human pancreatic tissues and the COVID-19 patients pancreatic tissues ( n = 10 images/group). Data are presented as mean ± SD. p values were calculated by unpaired two-tailed Student’s t test. *p < 0.05, **p < 0.01, and ***p < 0.001

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Infection, Activation Assay, Staining, Immunofluorescence, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet: SARS-CoV-2 infection triggers distinct characteristics of proteomics and metabolomics among adult, elder, and vaccinated COVID-19 NHP models. a Bar and dot plots of the KEGG pathway combined functional analysis of the enriched pathways associated with differentially expressed proteins and metabolites between the control and adult prototypic SARS-CoV-2 strain-infected model groups. b , c Heatmap revealing that genes corresponding to differential proteins and metabolites are mainly enriched in the insulin resistance pathway and lipid and atherosclerosis pathway. d Bar plots of the KEGG pathway combined functional analysis of the enriched pathways associated with differentially expressed proteins and metabolites from 3 days post-infection (DPI) to 7 DPI from the elder prototypic SARS-CoV-2 strain-infected COVID-19 model compared with those of the control group. In a and d , the blue characters represent the signaling pathway directly related to glycometabolism involved in the islets, the red characters represent the metabolic pathway directly related to COVID-19, and the green characters represent the metabolic pathway related to blood circulation. e Correlation networks of significant protein interaction. Each node in the interaction network represents a protein, and the change in node color from red to blue represents the change in protein expression level from being upregulated to being downregulated. The thickness of the lines represents the change from high to low confidence in interaction relationships. f Volcano plot showing the relative content differences of proteins between the control and the prototypic SARS-CoV-2 strain-infected model vaccinated with inactivated vaccines and the significance of statistical differences. Each dot in the volcano plot represents a protein, with green dots representing downregulated differential proteins, red dots representing upregulated differential proteins, and gray dots representing proteins detected but not significantly different. g , h Volcano plot showing the relative content differences of lipids and metabolites between the control and vaccinated COVID-19 model groups and the significance of statistical differences, respectively. Each dot in the volcano plot represents a lipid or a metabolite, with green dots representing downregulated differential lipids or metabolites, red dots representing upregulated differential lipids or metabolites, and gray dots representing lipids or metabolites detected but not significantly different. i Dynamic distribution of metabolites. The abscissa represents the cumulative number of substances in the order of difference multiple from small to large, and the ordinate represents the pair value with the difference multiple as base 2. Each dot represents a substance, the green dot represents the substance that is in the top 10 ranking down, and the red dot represents the substance that is in the top 10 ranking up. The KEGG pathway name with blue characters represents significantly enriched pathways related to diabetes mellitus and glucose metabolism impairment. The KEGG pathway name with red characters represents a SARS-CoV-2-associated pathway. The KEGG pathway name with green characters represents significantly enriched pathways related to thrombogenesis

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Infection, Functional Assay, Expressing, Vaccines

Journal: Signal Transduction and Targeted Therapy

Article Title: Infection with SARS-CoV-2 can cause pancreatic impairment

doi: 10.1038/s41392-024-01796-2

Figure Lengend Snippet:

Article Snippet: GeneTex:SARS-CoV-2 (COVID-19) nucleocapsid antibody [HL5511] , GeneTex , Cat# GTX635689.

Techniques: Marker, Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Glucose Assay, Polymer, Titration, Modification, Staining, Software