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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient <t>of</t> <t>G-actin</t> polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.
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Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient of G-actin polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.

Journal: Redox Biology

Article Title: Myo1f regulates monocyte adhesion and contributes to atherosclerosis via MRTFA-dependent ITGB2 expression

doi: 10.1016/j.redox.2026.104049

Figure Lengend Snippet: Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient of G-actin polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.

Article Snippet: The F-actin to G-actin ratio was determined following the manufacturer's instructions (BK037, Cytoskeleton).

Techniques: Translocation Assay, Western Blot, Expressing, Fluorescence, Microscopy, Immunofluorescence, Labeling, Staining, Knockdown, Binding Assay, Two Tailed Test