Journal: bioRxiv

Article Title: Virus Entry is a Major Determinant of HCMV Latency in Monocytes

doi: 10.1101/2024.10.26.619803

Figure Lengend Snippet: (a). Flow cytometry analysis of cell surface staining of PDGFRα versus IgG control in primary and THP1 monocytes and macrophages. (b). Flow cytometry analysis of cell surface staining of PDGFRα of THP1 and THP1- PDGFRα monocytes. (c). Light microscopy of THP1 and THP1-PDGFRα monocytes. (d). Differential expression analysis of RNA-seq from THP1 and THP1-PDGFRα monocytes. Blue dots mark significant differentially expressed genes ( p < 0.05, FC > 1).

Article Snippet: FITC-conjugated Mouse IgG1 anti-human CD29 (ITGB1) (Santa cruz catalog no. MEM-101A ) and FITC-conjugated Mouse IgG1 control (Santa Cruz, catalog no. sc-2339).

Techniques: Flow Cytometry, Staining, Control, Light Microscopy, Expressing, RNA Sequencing Assay

Journal: bioRxiv

Article Title: Virus Entry is a Major Determinant of HCMV Latency in Monocytes

doi: 10.1101/2024.10.26.619803

Figure Lengend Snippet: (a). NRP2 expression in primary, THP-1 and Kasumi-3 monocytes and macrophages as measured by RNA-seq. Mono, monocytes; mac, macrophages; RPM, reads per million. (b). Flow cytometry analysis of NRP2 versus IgG control Cell surface staining in primary and THP1 monocytes and macrophages. (c). Flow cytometry analysis of THP1 macrophages, transfected with NRP2 and control siRNA two days before infection with HCMV-GFP. Analysis performed at 3 d.p.i. Quantification of the replicates is presented on the right bar graph. (d). Flow cytometry analysis of infected THP1 macrophages with NRP2 or control CRISPR knockout using HCMV-GFP virus. Analysis performed at 3 d.p.i. Quantification of the replicates is presented on the right bar graph.

Article Snippet: FITC-conjugated Mouse IgG1 anti-human CD29 (ITGB1) (Santa cruz catalog no. MEM-101A ) and FITC-conjugated Mouse IgG1 control (Santa Cruz, catalog no. sc-2339).

Techniques: Expressing, RNA Sequencing Assay, Flow Cytometry, Control, Staining, Transfection, Infection, CRISPR, Knock-Out, Virus

Journal: bioRxiv

Article Title: Virus Entry is a Major Determinant of HCMV Latency in Monocytes

doi: 10.1101/2024.10.26.619803

Figure Lengend Snippet: (a). Relative expression level of NRP2, measured by real-time PCR in THP1 macrophages treated with NRP2 siRNA compared to control siRNA at 0 h.p.i. (b) Cell surface staining with NRP2 or IgG control of THP1 macrophages treated with CRISPR knockout against NRP2 or control. Cells were analyzed by Flow cytometry.

Article Snippet: FITC-conjugated Mouse IgG1 anti-human CD29 (ITGB1) (Santa cruz catalog no. MEM-101A ) and FITC-conjugated Mouse IgG1 control (Santa Cruz, catalog no. sc-2339).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Staining, CRISPR, Knock-Out, Flow Cytometry

Journal: bioRxiv

Article Title: Virus Entry is a Major Determinant of HCMV Latency in Monocytes

doi: 10.1101/2024.10.26.619803

Figure Lengend Snippet: (a). ITGB3 and ITGB1 expression in primary, THP1 and Kasumi-3 monocytes and macrophages as measured by RNA-seq. RPM, reads per million. (b-c). Flow cytometry analysis of integrin β1 (b) and integrin β3 (c) cell surface levels in primary and THP1 monocytes and macrophages. (d). Flow cytometry analysis of control, ITGB3, ITGB1 and ITGB3 + ITGB1 knockout (KO) in THP1 macrophages infected with HCMV–GFP. Cells were analyzed at 3 d.p.i. (e). Quantification of the replicates of . (f-g). Representative Microscopy images (f) and quantification using FIJI analysis software (g) of THP1 macrophages with ITGB3 knockout (KO) versus control knockout (n=60 in the control and n=67 in the ITGB3 KO). Cells were infected with HCMV-UL32-GFP and imaged at 1.h.pi. Actin staining was used to visualize the cell’s borders and DAPI for nuclei staining. (h). Flow cytometry analysis of infected THP1 monocytes overexpressing ITGB3 and ITGAV (αvβ3), compared to mCherry control. Overexpression was induced for 24 hours using doxycycline prior to infection. Cells were analyzed at 3 d.p.i. (i). Quantification of the replicates of . (j). Expression levels of ITGB3, ITGB1 and NRP2 in CD34+ HSCs from RNA-seq of eight healthy patients . RPKM, Reads Per Kilobase per Million mapped reads.

Article Snippet: FITC-conjugated Mouse IgG1 anti-human CD29 (ITGB1) (Santa cruz catalog no. MEM-101A ) and FITC-conjugated Mouse IgG1 control (Santa Cruz, catalog no. sc-2339).

Techniques: Expressing, RNA Sequencing Assay, Flow Cytometry, Control, Knock-Out, Infection, Microscopy, Software, Staining, Over Expression

Journal: bioRxiv

Article Title: Virus Entry is a Major Determinant of HCMV Latency in Monocytes

doi: 10.1101/2024.10.26.619803

Figure Lengend Snippet: (a). Cell surface staining of THP1 macrophages with a CRISPR knockout of β3 (left), β1 (right), or both versus control knockout. Cells were analyzed by Flow cytometry. (b). real-time PCR analysis of THP1 macrophages treated with siRNA against ITGB3 or control at 0 h.p.i.. (c). Flow cytometry analysis of THP1 macrophages, transfected with ITGB3 or control siRNAs and infected with HCMV–GFP. Cells were analyzed at 3 d.p.i. (d). Quantification of the FACS replicates of . (e). Cell surface staining of THP1 overexpressing (OE) αVβ3 compared to IgG control. Cells were stained for both αV and β3 in the αVβ3 overexpressing cells. Double positive cells are gated in a box.

Article Snippet: FITC-conjugated Mouse IgG1 anti-human CD29 (ITGB1) (Santa cruz catalog no. MEM-101A ) and FITC-conjugated Mouse IgG1 control (Santa Cruz, catalog no. sc-2339).

Techniques: Staining, CRISPR, Knock-Out, Control, Flow Cytometry, Real-time Polymerase Chain Reaction, Transfection, Infection