Journal: Life

Article Title: Characterization of ACE Inhibitors and AT 1 R Antagonists with Regard to Their Effect on ACE2 Expression and Infection with SARS-CoV-2 Using a Caco-2 Cell Model

doi: 10.3390/life11080810

Figure Lengend Snippet: Effect of ACE-inhibitors and AT 1 R antagonists on ACE2 mRNA expression. Caco-2 cells were incubated with ACE-inhibitors, AT 1 R antagonists, or vehicle in the indicated concentrations and time points. The mRNA expression was determined using quantitative PCR, normalized to β-actin and drug-treated samples were related to the vehicle treated samples (control) to obtain the fold induction. The experiment was achieved in three biological and three technical replicates. The means of three technical replicates are shown and used for statistical analysis (two-way ANOVA with Dunnett’s multiple comparisons test). * p < 0.01 show statistically significant difference between drug-treated and vehicle-treated samples.

Article Snippet: The mouse monoclonal anti-β-actin antibody was purchased from Sigma Aldrich (Schnelldorf, Germany), the rabbit polyclonal anti-ACE2 antibody from Abcam (Berlin, Germany), and the anti-rabbit AF488 and anti-mouse AF546 antibodies from Thermo Scientific (Schwerte, Germany).

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Control

Journal: Life

Article Title: Characterization of ACE Inhibitors and AT 1 R Antagonists with Regard to Their Effect on ACE2 Expression and Infection with SARS-CoV-2 Using a Caco-2 Cell Model

doi: 10.3390/life11080810

Figure Lengend Snippet: Effect of ACE-inhibitors and AT 1 R antagonists on ACE2 protein expression. Caco-2 cells were incubated with ACE-inhibitors, AT 1 R antagonists, or vehicle in the indicated concentrations and time points. The protein expression was determined using Western blot technology, normalized to β-actin and drug-treated samples were related to non-treated samples to obtain the fold induction. The experiment was achieved in three biological replicates. Two-way ANOVA with multiple comparisons test was used to analyse statistical difference between drug-treated and vehicle-treated samples.

Article Snippet: The mouse monoclonal anti-β-actin antibody was purchased from Sigma Aldrich (Schnelldorf, Germany), the rabbit polyclonal anti-ACE2 antibody from Abcam (Berlin, Germany), and the anti-rabbit AF488 and anti-mouse AF546 antibodies from Thermo Scientific (Schwerte, Germany).

Techniques: Expressing, Incubation, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: SARS-CoV-2 Spike Protein Induces Degradation of Junctional Proteins That Maintain Endothelial Barrier Integrity

doi: 10.3389/fcvm.2021.687783

Figure Lengend Snippet: Spike increases Rab5a association with ACE2 and junctional proteins. (A) Representative Western blot of coimmunoprecipitation experiments done using Rab5a antibody and probed for different proteins. (B) Mean data indicating fold change in Rab5a association with junctional proteins and ACE2 after S1RBD treatment. n = 6 for each, * P < 0.05 vs. untreated control. (C) Representative Western blot of coimmunoprecipitation experiments done using ACE2 antibody and probed for Rab5a. (D) Mean data indicating fold change in ACE2 association with Rab5a after Spike treatment. n = 6 for each, * P < 0.05 vs. untreated control. (E) Representative Western blot of coimmunoprecipitation experiments in control and diabetic cells after Rab5a knockdown with siRNA. (F) Mean data indicating fold change in ACE2 association with Rab5a after Rab5a siRNA. n = 6 for each, * P < 0.05 vs. control, # P < 0.05 vs. NT siRNA diabetic.

Article Snippet: Membranes were blocked with 5% non-fat milk and incubated with the one of the following primary antibodies: VE-Cadherin, PECAM1 (Novus Biologics), JAM-A (Invitrogen), Connexin-43, Rab5a (Cell Signaling Inc.), ACE2 (Abnova), or Actin (Millipore Sigma) overnight at 4°C.

Techniques: Western Blot, Control, Knockdown

Journal: Cell Reports

Article Title: Limited extent and consequences of pancreatic SARS-CoV-2 infection

doi: 10.1016/j.celrep.2022.110508

Figure Lengend Snippet: Stringent ACE2 requirement for pancreatic islet cell infection with SARS-CoV-2 (A) Representative contour plots gated on live α, β, and “other” cells pre-treated with IgG (irrelevant polyclonal goat antibody AF7197) or the anti-ACE2 blocking antibody AF933 prior to SARS-CoV-2 infection (48 h). (B) Summary of SARS-CoV-2 NP expression by live islet cell subsets as a function of IgG treatment or ACE2 blockade (n = 6 donors). (C) Percent infection inhibition for β and “other” cells (inhibition for α cells is not shown because the very low extent of α cell infection in IgG-treated cultures for 2 of 6 donors substantially skews such calculations). (D) Infectious SARS-CoV-2 titers and extent of infection inhibition following ACE2 blockade (n = 3 donors). (E) Quantification of chemokines and cytokines in UV-inactivated TCS of SARS-CoV-2-infected islet cell cultures under conditions of IgG treatment or ACE2 blockade (48-h infection, n = 3 donors). (F) Infectious SARS-CoV-2 titers in TCS as a function of glucose concentration in islet culture medium (n = 3 donors). (G) Quantification of CXCL10 and CXCL11 in TCS as a function of glucose concentration. All summary bar diagrams represent mean ± SD and scatter for the indicated number of donors; statistical analyses were conducted by paired t test or repeated-measures ANOVA with Tukey’s multiple comparisons where applicable (∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001). All summary bar diagrams represent mean ± SD and scatter for the indicated number of donors; statistical analyses were conducted by paired t test or repeated-measures ANOVA with Tukey’s multiple comparisons where applicable.

Article Snippet: Anti-human ACE2 (polyclonal goat IgG Poly5036) - AF647 (in-house conjugation) , Biolegend , Cat# 503602; RRID: AB_2892475.

Techniques: Infection, Blocking Assay, Expressing, Inhibition, Concentration Assay

Journal: Cell Reports

Article Title: Limited extent and consequences of pancreatic SARS-CoV-2 infection

doi: 10.1016/j.celrep.2022.110508

Figure Lengend Snippet:

Article Snippet: Anti-human ACE2 (polyclonal goat IgG Poly5036) - AF647 (in-house conjugation) , Biolegend , Cat# 503602; RRID: AB_2892475.

Techniques: Conjugation Assay, Purification, Blocking Assay, Recombinant, Control, Virus, Saline, Modification, Staining, Library Quantification, Antibody Labeling, Flow Cytometry, Software, Cytometry, Sequencing