Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Dysregulated expression of primary cilium-associated genes in SARS-CoV-2 infected patient lung samples. A The Venn diagram presents genome-wide RNA sequencing results from patient lung samples, revealing differentially expressed genes (DEGs) associated with primary cilium. The DEGs were defined by log2 fold change > 1.5, with adjusted P < 0.05. P values were adjusted using the Benjamini–Hochberg correction. The analysis showing 3,972 DEGs in patient samples and 956 primary cilia-related genes in CiliaCarta, with 129 overlapping genes functionally linked to primary cilium. B The list of 129 ciliary DEGs identified in the SARS-CoV-2 patient lung samples. Gene ranked by the differently expressed level. Blue dots represent down-regulated genes, red dots represent up-regulated genes. Colour gradient and size of dots correlate with the degree of fold change. C Volcano plots of 129 DEGs in the patient lung samples overlap with the genes associated with ciliary functions. DEGs at -log10 > 2 and |log2 fold change|> 2 were indicated. (adjusted p values < 0.05, P values adjusted using the Benjamini–Hochberg correction) 61 up-regulated genes presented as red dots, 68 down-regulated genes presented as blue dots. Top 15 and two prominent ciliary genes ARL13B and ADCY3 are indicated. D Gene Ontology (GO) enrichment analysis for Biological Processes among ciliary DEGs from SARS-CoV-2 patient lung samples. Terms are ranked by their enrichment score, calculated using Gene Set Enrichment Analysis (GSEA). E Gene Set Enrichment Analysis (GSEA) reveals key signaling pathways associated with ciliary DEGs from SARS-CoV-2 patient lungs, ranked by enrichment score. F Ciliary DEGs from SARS-CoV-2 patient lung samples grouped by ciliary functional category. G Venn diagram and table show overlap between ACE2 interactors from BioGRID (979 proteins) and ciliary genes from CiliaCarta (956 genes) and Syscilia (303 genes). 56 and 14 overlapping genes were identified in each cilia database, respectively. Two well-studied ciliary components, namely ARL13B and TULP3 are identified in the intersections (highlighted yellow). H Gene Ontology (GO) enrichment analysis for Biological Processes among ciliary ACE2 interactors. Terms are ranked by their enrichment score, calculated using Gene Set Enrichment Analysis (GSEA)

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques: Expressing, Infection, Genome Wide, RNA Sequencing, Protein-Protein interactions, Functional Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: TULP3 colocalizes with ACE2 on the primary cilium axoneme and physically interacts with ACE2. A Representative immunofluorescence images show the accumulation and co-localization of TULP3 and ACE2 on the primary cilium of A549 cells. B Histogram illustrates co-localization profile of endogenous ACE2 and TULP3 expressions on ARL13B-positive cilia axoneme of A549. C Mander’s Coefficients analysis of ACE2 and TULP3 co-localization in cytoplasm versus primary cilia of A549 cell. The data represents quantifications from three independent experiments, with 10–20 data points measured in each experiment. Statistical analysis is performed using Paired Student’s t-Test, Two tailed. P values: ** < 0.01, *** < 0.001, **** < 0.0001. D Representative Western-blot images illustrate the co-IP experiment of ACE2 and TULP3 on ACE2-GFP overexpressing HEK293T cell line. IP: immunoprecipitation; IB: immunoblot. E Graph depicts densitometry analysis of TULP3 signal intensity in ACE2 and IgG immunoprecipitation groups. The data represents quantifications from three independent experiments. Statistical analysis is performed using Unpaired Student’s t-Test, Two tailed. P value * < 0.05. F Graph depicts densitometry analysis of ACE2 signal intensity in TULP3 and IgG immunoprecipitation groups. The data represents quantifications from three independent experiments. Statistical analysis is performed using Unpaired Student’s t-Test, Two tailed. P value * < 0.05. G Representative Western-blot images illustrate the co-IP experiment of ACE2 and ARL13B on ACE2-GFP overexpressing HEK293T cell line. IP: immunoprecipitation; IB: immunoblot. H TULP3–ACE2 interaction interface predicted from AlphaFold3 . TULP3 (light blue, residues 271–430) and ACE2 (light red, residues 749–773) are shown with key interface residues highlighted as sticks (blue for TULP3, red for ACE2). The TULP3 cargo-binding face (β8–β12, highlighted in marine) engages the membrane-proximal ACE2 segment. Yellow dashed lines represent top recurring contacts across models. ACE2 residue I761, involved in all major contacts, is labelled

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Two Tailed Test, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation, Binding Assay, Membrane, Residue

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: TULP3 regulates ACE2 localization to the primary cilium. A Representative immunofluorescence images showing reduced ACE2 expression on the primary cilium in the shControl and TULP3 KD groups. B Graphs showing reduced ACE2 enrichment ratio in the (top) TULP3 and (bottom) ARL13B KD groups. ACE2 enrichment ratio was calculated as (ACE2 intensity per area on primary cilium)/(ACE2 intensity per area on whole cell). The data represents quantifications from three to four independent experiments, n = 14–17 measurements in each experiment. Statistical analysis is performed using (top) One-way ANOVA followed by Dunnett’s multiple comparisons and (bottom) Unpaired Student’s t-test. P values: * < 0.05, *** < 0.001, ns = not significant. C Representative immunofluorescence images show ciliary ACE2 expression on the primary cilium in TULP3 WT and TULP3 mut12 overexpression groups (pseudo-color magenta). D Graph shows ciliary ACE2 enrichment ratio in the TULP3 WT rescue and TULP3. mut12 overexpression groups. ACE2 enrichment ratio was calculated as (ACE2 intensity per area on primary cilium)/(ACE2 intensity per area on whole cell). The data represents quantifications from four independent experiments, n = 10–20 measurements in each experiment. Statistical analysis is performed using One-way ANOVA followed by Tukey’s multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Expressing, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Schematic diagram illustrating our proposed model. TULP3 functions as a ciliary adaptor protein that governs the targeting of ACE2 to the ciliary axoneme, acting partially through anchoring to the IFT-A transport complex

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Attachment and accumulation of the viral spike protein on the ACE2-enriched primary cilium. A Representative immunofluorescence images show the negative control without exposure to spike protein. A ” Histogram illustrating the ciliary co-localization and expression profiles on of spike and ACE2 signals the negative control. B Representative immunofluorescence images show the accumulation and co-localization of viral recombinant spike protein on endogenously expressed ACE2 on the ARL13B. + primary cilium. B ” Histogram illustrating the co-localization and expression profile of three fluorescence channels reflecting viral spike protein, ARL13B and ACE2 respectively, on the primary cilium. C Graph depicts the spike expression intensity on ACE2-negative and ACE2-positive primary cilia. ACE2 pixel intensities per cilia area greater than 500 were defined as ACE2-positive primary cilia, whereas below 500 were counted as ACE2-negative primary cilia. The data represent quantifications from three independent experiments, with n = 10–20 for each group in each experiment. P values: ** < 0.01

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Negative Control, Expressing, Recombinant, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Host cell entry factors for SARS-CoV-2 are endogenously expressed and enriched in primary cilia across multiple SARS-CoV-2 susceptible cell types. A Representative immunofluorescence images show the endogenous expression of ACE2 (green) on the primary cilium (red) of human lung carcinoma, A549, human retinal pigment epithelial cells (hTERT-RPE1), as well as neuronal cell types namely neuroblastoma, SH-SY5Y, and human induced-pluripotent stem cell derived neural progenitor cells (hiPSC-NPCs) respectively. B The violin plots depict the quantification of the per area expression intensity of ACE2 in the primary cilium in relation to the entire cell. The data represents quantifications from three independent experiments. Statistical analysis is performed using Wilcoxon matched-pairs signed rank test. P values: ** < 0.01,**** < 0.0001. ~ 10 to 25 cells were analysed in each group of three independent experiments. C The violin plot depicts the percentage of ACE2-expressing primary cilia across different human cell types that are susceptible to SARS-CoV-2 infection. Data illustrates three to eight independent experiments for each group. Statistical analysis is performed using Kruskal–Wallis Test and Dunn’s multiple comparisons test. P values: * < 0.05, ** < 0.01. D Representative immunofluorescence images showing the endogenous expression of NRP1 (green) on the primary cilium (red) of human lung carcinoma, A549 and hiPSC-NPCs

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Expressing, Derivative Assay, Infection

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Genetic perturbation of primary cilia formation reduces SARS-CoV-2 infection in human host cells. A - B Representative Western blot images and the corresponding graphs show the knockdown efficiency of sh ARL13B and sh IFT88 in the ( A ) A549 and ( B ) RPE1 knockdown cells. Knockdown efficiency was quantified by normalizing the signal intensity of the target protein from each knockdown sample to its respective control group. The data represents quantifications from three independent experiments. Statistical analysis is performed using One-way ANOVA followed by Dunnett’s multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ns = not significant. Error bar: SD. C Representative immunofluorescence images show a reduced number of ciliated cells in the ARL13B KD and IFT88 KD groups in A549 (top panel) and RPE1 (bottom panel) cell lines. D - E Graphs depict the percentage of primary cilia-bearing (ciliated) cells in the ARL13B KD and IFT88 KD groups in ( D ) A549 and ( E ) RPE1 respectively. The data represents quantifications from five independent experiments. Statistical analysis is performed using One-way ANOVA followed by Dunnett’s multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ns = not significant. F - G Graphs depict flow cytometry analysis of the relative infection rate of different SARS-CoV-2-GFP pseudovirus variants upon perturbation of primary cilia in the ARL13B - and IFT88- knockdown human lung and retinal cells respectively. The percentage of infected cells was determined by flow cytometry analysis of GFP-positive infected cells. The data represents quantifications from three independent experiments. Each dot represents the mean value of an independent experiment. Statistical analysis is performed using Two-way ANOVA, Tukey's multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ns = not significant. Error bar: S.E.M. H Schematic diagram summarizes the findings, showing that the loss of ACE2-enriched cilia decreases SARS-CoV-2 viral uptake. Created with https://BioRender.com

Article Snippet: The primary antibodies used include, anti-ACE2 antibody (1:3000, cat. number: AF933, R&D systems), anti-ARL13B antibody (1:3000, cat. number: 17711–1-AP, Proteintech), anti-IFT88 antibody (1:2000, cat. number: ab42497, Abcam), anti-TULP3 antibody (1:500, cat. number: 13637–1-AP, Proteintech), anti-KIF3A antibody (1:4000, cat. number: ab11259, Abcam), anti-NRP1 antibody (1:1000, cat. number: ab81321, Abcam), anti-GAPDH antibody (1:8000, cat. number: ab8245, Abcam), anti-ß-ACTIN antibody (1:2000, cat. number: sc-47778, Santa Cruz Biotechnology).

Techniques: Infection, Western Blot, Knockdown, Control, Immunofluorescence, Flow Cytometry

Journal: Research in Veterinary Science

Article Title: Elevated angiotensin-converting enzyme 2 (ACE2) expression in cats with hypertrophic cardiomyopathy

doi: 10.1016/j.rvsc.2022.09.024

Figure Lengend Snippet: ACE2 immunohistochemical analysis of the cat lung, heart, and kidneys from hypertrophic cardiomyopathy (HCM) and non-HCM cases. (a) Total score of abundance and intensity of ACE2 immunolabelling, expressed as sum of bronchiole, alveoli, heart, and kidney scores for each animal, were compared between the two groups. (b, c) Semi-quantitative and qualitative assessment were made for the abundance or intensity of ACE2, respectively. Score as follow: abundance - 0 (no positive staining), 1 (0 to <25% positive cells), 2 (≥25% to <50%), 3 (≥50% to <75%) and 4 (≥75% to 100%); intensity 0 (negative), 1 (weak), 2 (moderate), and 3 (strong). Ten cases were reviewed for both HCM and non-HCM. Height of bars (a, b, c) represent the mean value and the error bars represent standard deviation. Mann-Whitney test. * p < 0.05.

Article Snippet: Briefly, this involved antigen retrieval of FFPE sections in pH 6 buffer, immunolabelling with rabbit polyclonal antibody against ACE2 (Abcam) and Envision flex (Dako) and visualisation with DAB chromogen.

Techniques: Immunohistochemical staining, Staining, Standard Deviation, MANN-WHITNEY

Journal: Research in Veterinary Science

Article Title: Elevated angiotensin-converting enzyme 2 (ACE2) expression in cats with hypertrophic cardiomyopathy

doi: 10.1016/j.rvsc.2022.09.024

Figure Lengend Snippet: Immunohistochemical labelling of ACE2 in the heart and lung of cats from non-HCM (a, c) and HCM (b, d) cases. Low levels of ACE2 immunolabelling in the heart and lung of non-HCM cats, limited to the vascular endothelium (arrow head) and type I pneumocytes (arrow), respectively (a, c). ACE2 immunolabelling was more abundant and intense in the vasculature (arrowhead) within the heart (b), and more abundant and intense in the alveoli typically in the type I pneumocytes (arrow) and occasionally in type II pneumocytes (arrowhead) of cats with HCM (d). Images taken with 200× objectives.

Article Snippet: Briefly, this involved antigen retrieval of FFPE sections in pH 6 buffer, immunolabelling with rabbit polyclonal antibody against ACE2 (Abcam) and Envision flex (Dako) and visualisation with DAB chromogen.

Techniques: Immunohistochemical staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2 transfection increased the mRNA level of ACE2 and the protein levels of VEGF and IGF. (a) qRT-PCR analysis of the ACE2 mRNA level in the three types of CM. (b, c) ELISA analysis of VEGF and IGF protein levels in the three types of CM. ∗ P < 0.05 vs. EPC-CM; # P < 0.05 vs. Null-EPC-CM. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques: Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: Characterization of Ang II-induced EC injury model. (a) EC apoptosis. (b, c) Tube formation and migration abilities. (d) ACE2 mRNA level in ECs. (e) Representative JC-1 staining images and summary data of Δψ M in ECs. Δψ M: mitochondrion membrane potential (MMP, red/green). Bar: 50 μ m. (f) Summary data showing the percentage of cells with fragmented mitochondria. ∗ P < 0.05 vs. control. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques: Migration, Staining, Membrane, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2-EPC-CM decreased Ang II-induced EC apoptosis and dysfunction. (a) Representative flow plots of EC apoptosis. (b) Summarized data of EC apoptosis. (c) Tube formation. (d) Migration. ∗ P < 0.05 vs. vehicle; + P < 0.05 vs. Null-EPC-CM. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques: Migration

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2-EPC-CM upregulated ACE2 level and improved Ang II/Ang (1–7) balance on Ang II-induced ECs. (a) ACE2 mRNA level of ECs. (b) The ratio of Ang II/Ang (1–7). ∗ P < 0.05 vs. vehicle; + P < 0.05 vs. Null-EPC-CM; # P < 0.05 vs. ACE2-EPC-CM. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2-EPC-CM increased MMP and ATP production. (a) The change of MMP in ECs after coincubation with different CM. (b) The ATP levels in ECs after different treatments. ∗ P < 0.05 vs. vehicle; + P < 0.05 vs. Null-EPC-CM; # P < 0.05 vs. ACE2-EPC-CM. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2-EPC-CM decreased mitochondrion fragmentation. (a) Summary data showing the percentage of cells with fragmented mitochondria. (b) Representative confocal images showing the morphology of the mitochondrion. Enlarged images are magnifications of the mitochondria at the indicated area. Bar: 10 μ m. ∗ P < 0.05 vs. vehicle; + P < 0.05 vs. Null-EPC-CM; # P < 0.05 vs. ACE2-EPC-CM. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2-EPC-CM decreased the expressions of Nox2 and Nox4. (a, b) Representative bands and summarized data showing the expressions of Nox2 and Nox4 in ECs exposed to Ang II. ∗ P < 0.05 vs. vehicle; + P < 0.05 vs. Null-EPC-CM; # P < 0.05 vs. ACE2-EPC-CM. N = 6/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: The dysfunction of ECs from R+ mice with decreased levels of ACE2 expression and activity. (a) ACE2 mRNA level of ECs. (b) ACE2 protein level of ECs. (c) ACE2 activity of ECs. (d) EC apoptotic rate is increased in R+ mice. (e, f) EC function is impaired in R+ mice. ∗ P < 0.05 vs. vehicle; + P < 0.05 vs. WT. N = 10/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques: Expressing, Activity Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Exosome-Mediated Transfer of ACE2 (Angiotensin-Converting Enzyme 2) from Endothelial Progenitor Cells Promotes Survival and Function of Endothelial Cell

doi: 10.1155/2020/4213541

Figure Lengend Snippet: ACE2-EPC-EXs decreased apoptosis and improved the function of ECs from R+ mice. (a) The effects of ACE2-EPC-EXs on EC apoptosis. (b) The effects of ACE2-EPC-EXs on EC tube formation. (c) The effects of ACE2-EPC-EXs on EC migration. ∗ P < 0.05 vs. Veh; + P < 0.05 vs. EPCs; # P < 0.05 vs. EPC-EXs; & P < 0.05 vs. ACE2-EPC-EXs. N = 10/group.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 hr at room temperature and incubated with primary antibody against ACE2 (1:1000, Cell Signaling Technology), Nox2 and Nox4 (1 : 1000, Abcam) or β -actin (1 : 4000; Sigma, St. Louis, MO) at 4°C overnight.

Techniques: Migration

Journal: Biomedicines

Article Title: Pan-Cancer Analysis Identifies SNORA12 as a Prognostic Biomarker and Demonstrates Its Role in Upregulating TIGIT in Osteosarcoma

doi: 10.3390/biomedicines14030723

Figure Lengend Snippet: SNORA12 drives upregulation of the immune checkpoint TIGIT in osteosarcoma. ( A – D ) Representative Western blot analyses of TIGIT protein expression in the indicated cell lines under different treatments. SW1353 ( A ) and U2OS ( B ) osteosarcoma cells, as well as primary NK cells ( C ), were subjected to SNORA12 overexpression (OE-SNORA12), with respective controls (Control and OE-NC). NK92 cells ( D ), which endogenously express high levels of SNORA12, were subjected to SNORA12 knockdown (sh-SNORA12), with controls (Control and sh-NC). β-actin served as the loading control. ( E – H ) Quantitative analysis of TIGIT protein levels normalized to β-actin from ( A – D ), respectively. Data are presented as mean ± SD. Statistical significance was determined by Student’s t -test (* p < 0.05, ** p < 0.01, ns = not significant).

Article Snippet: After blocking with 5% non-fat milk, membranes were incubated overnight at 4 °C with primary antibodies against TIGIT (1:1000, Proteintech, Cat# 66699-1-Ig, Rosemont, IL, USA) and β-actin (1:5000, serving as loading control).

Techniques: Western Blot, Expressing, Over Expression, Control, Knockdown

Journal: Journal of Neuroimmune Pharmacology

Article Title: SARS-CoV-2 Spike Protein Disrupts Blood–Brain Barrier Integrity via RhoA Activation

doi: 10.1007/s11481-021-10029-0

Figure Lengend Snippet: A Western blot of cells treated with DsiRNA targeting ACE2 (i), and relative intensity of ACE2 in DsiRNA and control cells normalized to beta-actin (ii). B Permeability coefficients measured in channels seeded with ACE2-Knockdown (KD) HCMEC/D3 cells. *indicates p < 0.05 compared to control. n = 3 for all experiments. C− D Fluorescent images of vessels stained with DAPI (blue), Phalloidin (green), ZO-1( magenta) (isolated in ii)for 3 conditions: siRNA control (C), and siRNA ACE2-KD (D)

Article Snippet: Gels were blocked in 5% normal donkey serum and 3% BSA for 30 min at room temperature followed by incubating overnight with primary antibodies for either angiotensin converting enzyme II (ACE2) (Santa Cruz) or zonula occludin-1 (ZO-1)(CST), a tight-junction scaffolding protein (Dejana ).

Techniques: Western Blot, Control, Permeability, Knockdown, Staining, Isolation

Journal: Journal of Neuroimmune Pharmacology

Article Title: SARS-CoV-2 Spike Protein Disrupts Blood–Brain Barrier Integrity via RhoA Activation

doi: 10.1007/s11481-021-10029-0

Figure Lengend Snippet: A Relative intensity (RQ) of RhoA activation in channels exposed to flow measured with ELISA. B Permeability measurement of vessels exposed to experimental conditions at 0-h and 3-h time points. *Indicates p < 0.05 compared to all conditions. C TEER values of vessels exposed to experimental conditions at 0-h,1.5 h and 3-h time points. * Indicates p < 0.05 compared to all 0-h and 1.5-h time points and ** indicates p < 0.05 compared all other conditions. n = 3 for all experiments. D Fluorescent images of vessels treated with C3 Transferase and S1 spike protein stained with DAPI (blue), Phalloidin (green), ACE2 (magenta), and ZO-1 ( red) (isolated in ii)

Article Snippet: Gels were blocked in 5% normal donkey serum and 3% BSA for 30 min at room temperature followed by incubating overnight with primary antibodies for either angiotensin converting enzyme II (ACE2) (Santa Cruz) or zonula occludin-1 (ZO-1)(CST), a tight-junction scaffolding protein (Dejana ).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Permeability, Staining, Isolation

Journal: bioRxiv

Article Title: SARS-CoV-2 and SARS-CoV differ in their cell tropism and drug sensitivity profiles

doi: 10.1101/2020.04.03.024257

Figure Lengend Snippet: Differentially conserved positions in the Spike protein. A) A sequence view of the DCPs present in the Spike protein, with insets showing the receptor binding domain and the two cleavage sites. B) The S interface with ACE2 (cyan). The ACE2 interface is shown in blue spheres, DCPs in red. C) The V404=K417 DCP. D) The R426=N439 DCP, the left image shows SARS-CoV S R426, the image on the right show the equivalen N439 in SARS-CoV-2 S. E) SARS-CoV residues associated with altering ACE2 affinity and the residues at these positions in SARS-CoV-2 S.

Article Snippet: To investigate whether ACE2 or DPP4 receptors are involved in SARS-CoV-2 internalisation and replication, Caco2 cells were pre-treated for 30 min at 37°C with goat antibody directed against the human ACE2 or DDP4 ectodomain (R&D Systems, Wiesbaden-Nordenstadt, Germany).

Techniques: Sequencing, Binding Assay

Journal: bioRxiv

Article Title: SARS-CoV-2 and SARS-CoV differ in their cell tropism and drug sensitivity profiles

doi: 10.1101/2020.04.03.024257

Figure Lengend Snippet: N A) Western blots indicating cellular ACE2 and TMPRSS2 protein levels. B) CPE formation in SARS-CoV and SARS-CoV-2 (MOI 0.01)-infected ACE2-negative 293 cells and 293 cells stably expressing ACE2 cells (293/ACE2) 48h post infection. C) Immunostaining for double-stranded RNA in SARS-CoV-2 and SARS-CoV (MOI 0.01)-infected 293/ACE2 cells 48h post infection. D) Quantification of virus genomes by qPCR in SARS-CoV-2 and SARS-CoV (MOI 0.01)-infected 293/ACE2 cells 48h post infection. E) Cytopathogenic effect (CPE) formation in SARS-CoV-2 and SARS-CoV (MOI 0.01)-infected Caco2 cells in the presence of antibodies directed against ACE2 or DPP4 (MERS-CoV receptor) 48h post infection.

Article Snippet: To investigate whether ACE2 or DPP4 receptors are involved in SARS-CoV-2 internalisation and replication, Caco2 cells were pre-treated for 30 min at 37°C with goat antibody directed against the human ACE2 or DDP4 ectodomain (R&D Systems, Wiesbaden-Nordenstadt, Germany).

Techniques: Western Blot, Infection, Stable Transfection, Expressing, Immunostaining, Virus