ccr2 antagonist rs504393  (Tocris)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Roles played by IL-8 in altering dynamics of trabecular meshwork cells after human cytomegalovirus infection

doi: 10.3389/fcimb.2025.1550509

Figure Lengend Snippet: Network analysis of the roles of CXCR2 and CCR2 in human trabecular meshwork cells (HTMCs) after HCMV infection. HCMV-infected HTMCs blocked by CXCR2 or CCR2 were analyzed using RNA-seq analysis. The significantly induced or repressed genes were assessed for canonical analysis using Ingenuity pathway analysis. Associations with detected canonical pathways were assessed by the z-scores. (A) The top repressed canonical pathways by CXCR2 blockade by SB225002 were the translocation of SLC2A4 (GLUT4) to the plasma membrane, remodeling of adherens junctions, Rho GTPases activated IQGAPs, aggrephagy, carboxyterminal post-translational modification of tubulin, viral replication pathway, Rho GTPases activates Formins, kinesins, and Gap junction trafficking and regulations (blue bars). (B) The repressed canonical pathways by CCR2 blockade by RS504393 were the cell cycle checkpoints and synthesis of DNA.

Article Snippet: For blocking CCR2, 5 μM RS504393 (CCR2 antagonist, Tocris, Bristol, UK) was used.

Techniques: Infection, RNA Sequencing, Translocation Assay, Clinical Proteomics, Membrane, Modification

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Roles played by IL-8 in altering dynamics of trabecular meshwork cells after human cytomegalovirus infection

doi: 10.3389/fcimb.2025.1550509

Figure Lengend Snippet: HCMV-induced cell motility of human trabecular meshwork cells (HTMCs). HTMCs infected with TB40/E were stained for actin using SiR-actin. (A) HCMV infected HTMCs were assessed for cell motility. HCMV infection significantly stimulated speed of cell movement. Exposure to SB225002 (500 nM) abolished HCMV-induced cell motility. The blockade of CCR2 by RS504393 (5 μM) partially reduced cell motility by HCMV infection. The data and images represent the speed of cell movement at 6 hours post-infection. For analysis, 4 three-dimensional frames of 1410 μm × 1060 μm × 90 μm were randomly selected from the wells by a masked observer, and cells whose movement could be traced within the same frame over 24 hours were extracted for digitization. Next, a randomly selected eight representative cells per group were analyzed by a masked observer for their speed of cell movement at each hourly interval using a multivariate linear regression analysis. P-values were adjusted using the Scheffe test. Data represent repeated experiments. (B) HCMV-infected HTMC were accompanied by an increased formation of lamellipodia (arrowhead) and filopodia (arrow). Exposure to SB225002 (500 nM) suppressed formation of lamellipodia and filopodia.

Article Snippet: For blocking CCR2, 5 μM RS504393 (CCR2 antagonist, Tocris, Bristol, UK) was used.

Techniques: Infection, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Roles played by IL-8 in altering dynamics of trabecular meshwork cells after human cytomegalovirus infection

doi: 10.3389/fcimb.2025.1550509

Figure Lengend Snippet: HCMV induced contraction of trabecular meshwork cells. HCMV-infected HTMCs were embedded in collagen gel, and assessed for cell contraction at 6 h. Cell contraction was calculated by the percentage of collagen gel area subtracted from total area of the well. HCMV (TB40/E) infection significantly induced cell contraction. Cell contraction was suppressed by blocking CXCR2 by SB225002 (500 nM). In contrast, cell contraction was not affected by RS504393 exposure. N=6/group. ANOVA and Tukey test. Data represent repeated experiments.

Article Snippet: For blocking CCR2, 5 μM RS504393 (CCR2 antagonist, Tocris, Bristol, UK) was used.

Techniques: Infection, Blocking Assay