Journal: Dyes and Pigments

Article Title: Potent inhibition of Severe Acute Respiratory Syndrome Coronavirus 2 by photosensitizers compounds

doi: 10.1016/j.dyepig.2021.109570

Figure Lengend Snippet: Screening of two photosensitizers by SARS-CoV-2 pseudoviruses entry assay. (A) Dose-responses of the inhibition of viral infectivity were also measured for photosensitizer ZnPc5K and ce6, giving the EC 50 and EC 90 . The photosensitizers completely inhibited the viral infectivity at 1 μM concentration upon LED illumination. The EGFP intensities of all fields in a microplate well at different concentrations were summed to represent the virus infected into the cells. Error bars represent the SD of duplicates in one experiment. Scale bar, 500 μm. (B) Photocytotoxicity of ZnPc5k and ce6 on hACE2-293T cells was measured by CCK-8 assay and showed almost no phototoxicity up to 5 μM with a light dose of 0.48 J/cm 2 . (C) Photosensitizers inactivated the SARS-CoV-2 pseudoviruses and reduced virus load inside ACE2-293T cells upon LED illumination. Serially diluted ZnPc5K or ce6 and pseudoviruses were incubated with ACE2-293T cells for 4 h, and illuminated by LED to a light dose of 0.48 J/cm 2 . After further 44 h incubation, the fluorescent images (green for EGFP for virus and blue for Hoechst for nucleus staining) were recorded and analyzed. Error bars represent the SD of triplicates in one experiment. Scale bar, 500 μm.

Article Snippet: The pLVX-hACE2 plasmid was constructed by cloning the coding region from Human ACE2 cDNA ORF Clone with N-terminal Flag tag (SinoBiological, HG10108-NF, China) into pLVX-IRES-Puro Lentiviral vector between Xho I and Not I sites.

Techniques: Inhibition, Infection, Concentration Assay, Virus, CCK-8 Assay, Incubation, Staining

Journal: bioRxiv

Article Title: Recombinant Human ACE2-Fc : A promising therapy for SARS-CoV2 infection

doi: 10.1101/2022.07.30.501940

Figure Lengend Snippet: a: Western blot analysis of MFcS2 and Ace2-Fc from conditioned supernatants from transient transfections with and without the plasmids in non-reducing condition b: SDS-PAGE analysis of purified MFcS2 and Ace2-Fc in reducing condition c: Western blot of purified Ace2-Fc in reduced condition d: Western blot of unreduced (UR) and reduced (R) MFcS2 protein purified from stably expressing HEK293E_MFcS2 cell line. The blots were probed with Goat anti-Human HRP and developed with DAB/H2O2

Article Snippet: ACE2 cDNA ORF clone (HG10108-ACR) was purchased from Sino biological, China.

Techniques: Western Blot, Transfection, SDS Page, Purification, Stable Transfection, Expressing

Journal: bioRxiv

Article Title: Recombinant Human ACE2-Fc : A promising therapy for SARS-CoV2 infection

doi: 10.1101/2022.07.30.501940

Figure Lengend Snippet: Binding affinity of MFcS2 and Ace2-Fc protein to the SARS-CoV2 Spike protein demonstrated by ELISA

Article Snippet: ACE2 cDNA ORF clone (HG10108-ACR) was purchased from Sino biological, China.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Recombinant Human ACE2-Fc : A promising therapy for SARS-CoV2 infection

doi: 10.1101/2022.07.30.501940

Figure Lengend Snippet: ACE2 and TEMPRSS2 expression in MhCT08-E cells cultured in low (11.1 mM) and high (25mM) glucose medium

Article Snippet: ACE2 cDNA ORF clone (HG10108-ACR) was purchased from Sino biological, China.

Techniques: Expressing, Cell Culture

Journal: Experimental & Molecular Medicine

Article Title: Generation of a lethal mouse model expressing human ACE2 and TMPRSS2 for SARS-CoV-2 infection and pathogenesis

doi: 10.1038/s12276-024-01197-z

Figure Lengend Snippet: a An experimental graphic created with BioRender.com. A549 cells or C57BL/6 mice were transfected with plasmid DNA as indicated to overexpress hACE2 and hTMPRSS2 in vitro and in vivo, respectively. b Plasmids expressing hACE2-FLAG (2 μg) or hTMPRSS2-HA (0.3 μg) were transfected into A549 cells on a 6-well plate. Overexpressed proteins at 24 h post transfection were analyzed using western blotting. GAPDH served as a loading control. c At 24 h post-transfection of plasmids, the cells were incubated with SARS-CoV-2-PV (1.3 × 10 5 TCID50/ml). After 24 h, the infectivity of the pseudovirus (PV) was assessed by measuring relative luminescence intensity. d At 2 days post-transfection, the cells were infected with SARS-CoV-2, followed by a further 2 days of incubation. The intracellular RNA was extracted and used to detect viral RNA of the SARS-CoV-2 nucleocapsid protein (NP) via RT-qPCR. e – g In vivo-transfected mice were intranasally infected with SARS-CoV-2 (5 × 10 5 PFU). Expression levels of hACE2 ( e ), hTMPRSS2 ( f ), and viral load ( g ) in the lungs were measured at the indicated days post-infection using RT-qPCR. Symbols represent the means ± SEM. Statistically significant differences between the groups were determined using one-way ANOVA ( c , d ) or unpaired two-tailed t -tests ( g ).

Article Snippet: Cells (2 × 10 5 cells per well) were plated into 6-well plates and transfected with 2 μg pCMV3-hACE2-FLAG (NM_021804.1, HG10108-CF; Sino Biological, Beijing, China) and/or 0.3 μg pCMV3-hTMPRSS2-HA (NM_005656.3, HG13070-CY; Sino Biological) using a TransIT-LT1 Transfection Reagent (Mirus Bio, Madison, WI, USA) according to the manufacturer’s recommendations.

Techniques: Transfection, Plasmid Preparation, In Vitro, In Vivo, Expressing, Western Blot, Incubation, Infection, Quantitative RT-PCR, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Generation of a lethal mouse model expressing human ACE2 and TMPRSS2 for SARS-CoV-2 infection and pathogenesis

doi: 10.1038/s12276-024-01197-z

Figure Lengend Snippet: a Experimental graphic for generating the double-Tg mice. Expression levels of hACE2 ( b ) and hTMPRSS2 ( c ) mRNA in the lungs, brain, heart, liver, kidney, and colon of the double-Tg or wild-type C57BL/6 mice (non-Tg) were assessed via RT-qPCR. Symbols represent the means ± SEM.

Article Snippet: Cells (2 × 10 5 cells per well) were plated into 6-well plates and transfected with 2 μg pCMV3-hACE2-FLAG (NM_021804.1, HG10108-CF; Sino Biological, Beijing, China) and/or 0.3 μg pCMV3-hTMPRSS2-HA (NM_005656.3, HG13070-CY; Sino Biological) using a TransIT-LT1 Transfection Reagent (Mirus Bio, Madison, WI, USA) according to the manufacturer’s recommendations.

Techniques: Expressing, Quantitative RT-PCR