Journal: Journal of medical virology

Article Title: SARS-CoV-2 pseudovirus infectivity and expression of viral entry-related factors ACE2, TMPRSS2, Kim-1 and NRP-1 in human cells from respiratory, urinary, digestive, reproductive and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: mRNA expression of ACE2, TMPRSS2, Kim‐1, and NRP‐1 in different types of cells from five human organ systems was analyzed by RT‐qPCR. (A−D) Relative mRNA expression levels of ACE2 (A), TMPRSS2 (B), Kim‐1 (C), and NRP‐1 (D) presented as relative C t values. β‐actin gene was used for normalization. Kim‐1, kidney injury molecule‐1; mRNA, messenger RNA; NRP‐1, neuropilin‐1; RT‐qPCR, real‐time quantitative polymerase chain reaction; TMPRSS2, transmembrane serine protease 2

Article Snippet: The primary antibodies used were a rabbit polyclonal antibody to ACE2 (ab15348, Abcam), rabbit monoclonal antibodies to TMPRSS2 (EPR3861) (ab92323, Abcam), Kim‐1 (E1R9N) (14971 S, CST) and NRP‐1 (EPR3113) (ab81321, Abcam), and a mouse monoclonal antibody to β‐tubulin.

Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Journal of medical virology

Article Title: SARS-CoV-2 pseudovirus infectivity and expression of viral entry-related factors ACE2, TMPRSS2, Kim-1 and NRP-1 in human cells from respiratory, urinary, digestive, reproductive and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: Blocking of infection by SARS‐CoV‐2 and SARS‐CoV pseudoviruses with S1, RBD, and other recombinant proteins of entry‐related factors in Huh‐7 cells. (A−E) Blocking of infection of SARS‐CoV‐2 and SARS‐CoV pseudoviruses by S1 (A), RBD (B), ACE2 (C), Kim‐1 (D), and NRP‐1 (E) recombinant proteins. ANOVA analysis was performed if multiple samples were involved. p‐value was calculated by unpaired two‐tailed Student's t test between uninfected cells and untreated infected cells, and between treated group and untreated infected group. p < 0.05 was considered as statistically significant. ANOVA, analysis of variance; RBD, receptor‐binding domain; Kim‐1, kidney injury molecule‐1; NRP‐1, neuropilin‐1; SARS‐CoV‐2, severe acute respiratory syndrome coronavirus 2; SARS‐CoV, severe acute respiratory syndrome coronavirus

Article Snippet: The primary antibodies used were a rabbit polyclonal antibody to ACE2 (ab15348, Abcam), rabbit monoclonal antibodies to TMPRSS2 (EPR3861) (ab92323, Abcam), Kim‐1 (E1R9N) (14971 S, CST) and NRP‐1 (EPR3113) (ab81321, Abcam), and a mouse monoclonal antibody to β‐tubulin.

Techniques: Blocking Assay, Infection, Recombinant, Two Tailed Test, Binding Assay

Journal: Journal of medical virology

Article Title: SARS-CoV-2 pseudovirus infectivity and expression of viral entry-related factors ACE2, TMPRSS2, Kim-1 and NRP-1 in human cells from respiratory, urinary, digestive, reproductive and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: Analyses of the correlation between the infectivity of SARS‐CoV‐2 or SARS‐CoV and the protein expression levels of entry‐related factors. (A–J) Correlation of the infectivity of SARS‐CoV‐2 (A, C, E, G, and I) or SARS‐CoV (B, D, F, H, and J) with glycosylated ACE2 (gly‐ACE2) (A and B), unglycosylated ACE2 (ungly‐ACE2) (C and D), TMPRSS2 (E and F), Kim‐1 (G and H) and NRP‐1 (I and J). The “red” dots represent cell lines/types that were infected by the pseudoviruses. The “green” dots represent cell lines/types that were not infected by the pseudoviruses. Kim‐1, kidney injury molecule‐1; NRP‐1, neuropilin‐1; SARS‐CoV‐2, severe acute respiratory syndrome coronavirus 2; SARS‐CoV, severe acute respiratory syndrome coronavirus; TMPRSS2, transmembrane serine protease 2

Article Snippet: The primary antibodies used were a rabbit polyclonal antibody to ACE2 (ab15348, Abcam), rabbit monoclonal antibodies to TMPRSS2 (EPR3861) (ab92323, Abcam), Kim‐1 (E1R9N) (14971 S, CST) and NRP‐1 (EPR3113) (ab81321, Abcam), and a mouse monoclonal antibody to β‐tubulin.

Techniques: Infection, Expressing

Journal: Journal of medical virology

Article Title: SARS-CoV-2 pseudovirus infectivity and expression of viral entry-related factors ACE2, TMPRSS2, Kim-1 and NRP-1 in human cells from respiratory, urinary, digestive, reproductive and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: Expression levels of ACE2, TMPRSS2, Kim‐1, and NRP‐1 proteins in different cell lines/types, and examination of ACE glycosylation in Huh‐7 cells analyzed by Western blot analysis. (A) Expression levels of ACE2, TMPRSS2, Kim‐1, and NRP‐1 proteins in different cell lines/types. Results showed that there are different expression profiles of ACE2, TMPRSS2, Kim‐1, and NRP‐1 in different cell lines/types. (B) Examination of ACE glycosylation in Huh‐7 cells. Untreated and PNGase F‐treated cell lysates were examined. β‐tubulin was used for loading normalization. Two bands of ~85 kD and ~120 kD were detected for ACE2 protein representing the unglycosylated and glycosylated forms of ACE2 (ungly‐ACE2 and gly‐ACE2), respectively. Kim‐1, kidney injury molecule‐1; NRP‐1, neuropilin‐1; TMPRSS2, transmembrane serine protease 2

Article Snippet: The primary antibodies used were a rabbit polyclonal antibody to ACE2 (ab15348, Abcam), rabbit monoclonal antibodies to TMPRSS2 (EPR3861) (ab92323, Abcam), Kim‐1 (E1R9N) (14971 S, CST) and NRP‐1 (EPR3113) (ab81321, Abcam), and a mouse monoclonal antibody to β‐tubulin.

Techniques: Expressing, Western Blot

Journal: Journal of medical virology

Article Title: SARS-CoV-2 pseudovirus infectivity and expression of viral entry-related factors ACE2, TMPRSS2, Kim-1 and NRP-1 in human cells from respiratory, urinary, digestive, reproductive and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: Expression levels and patterns of ACE2, Kim‐1, and NRP‐1 proteins in nine selected cell types/lines analyzed by IFA. IFA, Immunofluorescence assay; Kim‐1, kidney injury molecule‐1; NRP‐1, neuropilin‐1

Article Snippet: The primary antibodies used were a rabbit polyclonal antibody to ACE2 (ab15348, Abcam), rabbit monoclonal antibodies to TMPRSS2 (EPR3861) (ab92323, Abcam), Kim‐1 (E1R9N) (14971 S, CST) and NRP‐1 (EPR3113) (ab81321, Abcam), and a mouse monoclonal antibody to β‐tubulin.

Techniques: Expressing, Immunofluorescence

Journal: Nutrients

Article Title: Isoflavone Protects the Renal Tissue of Diabetic Ovariectomized Rats via PPARγ

doi: 10.3390/nu14132567

Figure Lengend Snippet: Isoflavone protects diabetic renal tissue of ovariectomized animals. Immunoreactivity of KIM-1 by immunohistochemistry ( a – f ). Control ( a ), OVX ( b ), DM1 ( c ), untreated DM1 + OVX ( d ), DM1 + OVX treated with isoflavone ( e ), and DM1 + OVX treated with 17β-estradiol ( f ). Densitometry analysis is represented in arbitrary units ( g ). Scale bar = 50 μm. Values represent mean ± SEM ( n = 6). Kruskal–Wallis test was performed, followed by Dunn’s multiple comparison test for non-normal distribution of data. ** p < 0.01; **** p < 0.0001.

Article Snippet: Paraffin sections were subjected to alcohol and xylene gradient solutions, antigen retrieval, protein block, and incubation with primary antibodies against PPAR-γ polyclonal antibody (1:200, Abcam, MA, USA) and kidney injury molecule 1 (KIM-1) (1:500, rabbit IgG, Sino Biological, Beijing, China) overnight at 4 °C.

Techniques: Immunohistochemistry