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: Hypertension

Article Title: Origin of the Y Chromosome Influences Intrarenal Vascular Responsiveness to Angiotensin I and Angiotensin (1-7) in Stroke-Prone Spontaneously Hypertensive Rats

doi: 10.1161/hypertensionaha.114.03756

Figure Lengend Snippet: Figure 2. Renal and renal artery mRNA gene expression of angiotensin-converting enzyme (ACE), ACE2, neprilysin, angiotensin II receptor type 1a (AT1aR), angiotensin II receptor type 2 (AT2R), and Mas receptor (MasR) in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n>5 per group). All relative gene expression data are expressed relative to the WKY group and are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.005 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.

Article Snippet: Taqman predesigned assays (Applied Biosystems) for ACE (Rn00561094_m1), ACE2 (Rn01416293_m1), angiotensin II receptor type 1a (Rn02758772_s1), angiotensin II receptor type 2 (AT2R, Rn00560677_s1), and Mas receptor (MasR, Rn00562673_s1) were multiplexed with Actb (4352340E-labeled VIC).

Techniques: Gene Expression

Journal: Hypertension

Article Title: Origin of the Y Chromosome Influences Intrarenal Vascular Responsiveness to Angiotensin I and Angiotensin (1-7) in Stroke-Prone Spontaneously Hypertensive Rats

doi: 10.1161/hypertensionaha.114.03756

Figure Lengend Snippet: Figure 3. Plasma angiotensin II (Ang II) and Ang (1-7) concentration, angiotensin-converting enzyme (ACE), and ACE2 activity in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n≥5/ strain). Data are presented as mean±SEM with *P<0.05, **P<0.01 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.

Article Snippet: Taqman predesigned assays (Applied Biosystems) for ACE (Rn00561094_m1), ACE2 (Rn01416293_m1), angiotensin II receptor type 1a (Rn02758772_s1), angiotensin II receptor type 2 (AT2R, Rn00560677_s1), and Mas receptor (MasR, Rn00562673_s1) were multiplexed with Actb (4352340E-labeled VIC).

Techniques: Clinical Proteomics, Concentration Assay, Activity Assay

Journal: Journal of Molecular and Cellular Cardiology

Article Title: Sex- and age-specific regulation of ACE2: Insights into severe COVID-19 susceptibility

doi: 10.1016/j.yjmcc.2021.11.003

Figure Lengend Snippet: Assessment of ACE2 and TMPRSS2 across sex and aging. A. ACE2 mRNA expression ( n = 36, biological replicates), protein levels ( n = 11, biological replicates), and activity (n = 36, biological replicates) are positively correlated across all organs in male and female animals (B . ). C. TMPRSS2 mRNA (n = 36, biological replicates) and protein levels (n = 11, biological replicates). Data from A-C was generated from C57BL6/J mice using a pooled analysis of males and females. Levels are compiled from all sexes and age groups for expression and activity; however, protein levels are compiled from representative and three replicate gels sampled from a subset of all age groups (Supplementary Fig. S1). All organs were run on the same gel for comparison. Immunoblots were over-exposed (O.E.) to visualize low levels in the lung and heart. Expression and activity data were run one time in the same plate for quantification. Data are represented as median with interquartile range (IQR) and were analyzed as independent samples with the Kruskal-Wallis test with pairwise comparison adjusted by Bonferroni correction; * indicates differences from the heart; # indicates differences from the lung; and † indicates differences from the kidney. * p < 0.05; ** p < 0.01, *** p < 0.001 where the number of symbols indicates the level of significance. Analysis of mRNA expression (D.), protein levels (E.), and ACE2 activity (F.) of ACE2 and TMPRSS2 across sex and age in the heart (magenta), lung (green), kidney (yellow), and small intestine (SI) (blue) ( n = 6, biological replicates/group). ACE2 global knockout mice were analyzed as a negative control (KO) for western blots. Raw images are provided in Supplementary Fig. S2-S3. Data are represented as the mean ± SEM and analyzed by two-way ANOVA with Tukey post hoc test (mRNA and activity), or unpaired student's t- test (protein levels). For all experiments, young animals ( n = 12) were obtained from four litters and each litter was collected over two days. Adult and aged animals (n = 12 /group) were obtained from five and three litters respectively, and each litter was collected over two days. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Real-time quantitative PCR was performed with TaqMan premixed assays (ThermoFisher Scientific) for gene expression in mice for TMPRSS2 (Mm00443687_m1) and ACE2 (Mm01159009_m1), and in humans for ACE2 (Hs00222343_m1).

Techniques: Expressing, Activity Assay, Generated, Comparison, Western Blot, Knock-Out, Negative Control

Journal: Journal of Molecular and Cellular Cardiology

Article Title: Sex- and age-specific regulation of ACE2: Insights into severe COVID-19 susceptibility

doi: 10.1016/j.yjmcc.2021.11.003

Figure Lengend Snippet: Sex- and age-differences in ACE2 levels in mouse and human hearts. A. Representative immunoblot of mouse hearts for ACE2 protein levels ( n = 6 biological replicates/group). Three gels were compiled for quantification (Supplementary Fig. S4). Data are represented as the mean ± SEM and were analyzed by two-way ANOVA with Tukey post hoc test; * p < 0.05; ** p < 0.01, *** p < 0.001. For all experiments, young animals ( n = 12) were obtained from four litters and each litter was collected over two days. Adult and aged animals ( n = 12 /group) were obtained from five and three litters respectively, and each litter was collected over two days. B. Immunofluorescence of ACE2 (green) and pericyte marker NG2 (red). Wheat Germ Agglutinin (WGA) (blue) was used to delineate the cell membrane. Qualitative images were captured from n = 2 biological replicates and n = 16 (8 images/animal) technical replicates for each age group. C. Representative immunoblot for ACE2 in human hearts ( n = 6, biological replicates/group). Two gels were compiled for quantification (Supplementary Fig. S4). Quantification represents the combined result from two western blots. Data are represented as the mean ± SEM and were analyzed by two-way ANOVA with Tukey post hoc test. * indicates differences from young males, and # indicates differences between aged males and aged females; * p < 0.05; ** p < 0.01, *** p < 0.001 where the number of symbols indicates the level of significance. D. Immunofluorescence of ACE2 (green) and NG2 (red). WGA (blue) was used to delineate the cell membrane. Qualitative images were captured from n = 2 biological replicates and n = 16 (8 images/donor) technical replicates for each age group. E. ACE2 protein levels and activity are positively correlated, but not mRNA expression. * indicates differences from young males, and # indicates differences between aged males and aged females. * p < 0.05; ** p < 0.01, *** p < 0.001 where the number of symbols indicates the level of significance. F. Schematic figure showing the multi-organ impact of sex and aging on ACE2 levels as a potential contributor to the increased male susceptibility to severe COVID-19. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Real-time quantitative PCR was performed with TaqMan premixed assays (ThermoFisher Scientific) for gene expression in mice for TMPRSS2 (Mm00443687_m1) and ACE2 (Mm01159009_m1), and in humans for ACE2 (Hs00222343_m1).

Techniques: Western Blot, Immunofluorescence, Marker, Membrane, Activity Assay, Expressing

Journal: Journal of Molecular and Cellular Cardiology

Article Title: Sex- and age-specific regulation of ACE2: Insights into severe COVID-19 susceptibility

doi: 10.1016/j.yjmcc.2021.11.003

Figure Lengend Snippet:

Article Snippet: Real-time quantitative PCR was performed with TaqMan premixed assays (ThermoFisher Scientific) for gene expression in mice for TMPRSS2 (Mm00443687_m1) and ACE2 (Mm01159009_m1), and in humans for ACE2 (Hs00222343_m1).

Techniques: