itga5  (Proteintech)

Journal: Advanced Science

Article Title: The Osteoblastic Microenvironment Determines the Fate of Breast Cancer Cells Disseminated in the Bone Marrow

doi: 10.1002/advs.202509980

Figure Lengend Snippet: RUNX2 increases the accumulation of MDA231 cells as micrometastases in bone marrow. (A) Experimental schedule for the bone colonization of MDA231‐derived cells in SCID mice. An osteogenic premetastatic niche (PMN) was established by injecting MDA231 CDH11 high /ITGA5 high extracellular vesicles into SCID mice via the tail vein for 3 weeks (2 doses/week). MDA231 RUNX2‐OE cells and control cells were injected into the mice via the left ventricle. (B) H&E staining images demonstrating osteoblasts (indicated by red triangles) in homeostatic bone (HB) and the PMN. (C) X‐ray images showing the bone mass in the HB and the PMN microenvironments. (D) Bar charts quantifying the osteoblast number as the ratio of osteoblast counts to bone perimeter in /mm (N.Ob/B.Pm) and bone mass as the bone volume fraction (BV/TV). (E) Western blot analysis demonstrating increased RUNX2 protein levels in MDA231 RUNX2‐OE cells compared with those in control cells. (F) Representative X‐ray images, pan‐cytokeratin (pan‐CK) immunohistochemical staining images, and TRAP staining images showing osteolytic lesions, tumor cell distribution, and activated osteoclasts, respectively. (G) Pie charts depicting the incidence of DTCs, micrometastases (micromets), and osteolytic lesions formed by control cells and RUNX2‐OE cells within HB and the PMN. The scale bars in the inset images indicate 20 µm. (H) Bar charts quantifying the tumor surface, erosion surface, and the number of TRAP + osteoclasts normalized to the total bone surface (N.Oc/BS in mm 2 ). (I) Bar charts illustrating the abundance and size of micrometastases in the bone marrow of mice without detectable bone lesions. (J) Representative KI67 fluorescence immunohistochemical staining images. Pan‐CK was used to label the tumor cells, while DAPI was used to stain the nuclei. (K) Bar chart illustrating the reduced numbers of KI67 + tumor cells in micrometastases compared with tumor cells within bone colonization. The data are displayed as the means ± SDs. n.s., not significant; * p <0.05, ** p <0.01, *** p <0.001 compared with the corresponding controls, as determined by Student's t‐test.

Article Snippet: Additionally, ATN‐161 (1 mg/kg, MCE, HY‐13535A) was employed to disrupt the tumor–bone matrix interaction mediated by ITGA5.

Techniques: Derivative Assay, Control, Injection, Staining, Western Blot, Immunohistochemical staining, Fluorescence

Journal: Advanced Science

Article Title: The Osteoblastic Microenvironment Determines the Fate of Breast Cancer Cells Disseminated in the Bone Marrow

doi: 10.1002/advs.202509980

Figure Lengend Snippet: The ITGA5 antagonist ATN‐161 reduces the incidence of bone colonization. (A–D) Experimental diagram (A), representative X‐ray, H&E staining, TRAP staining, and KI67 and pan‐CK immunofluorescence staining images (B), and bar charts demonstrating the inhibition of PTH‐reactivated bone colonization (C) and tumor surface normalized to the bone surface (D), in MDA231‐derived cells following ATN‐161 administration. (E–H) Experimental diagram (E), representative X‐ray, H&E staining, TRAP staining, and KI67 and pan‐CK immunofluorescence staining images (F), and bar charts displaying the blockade of E2‐supported bone colonization (G) and tumor surface normalized to the bone surface (H), in MCF7‐derived cells after ATN‐161 administration. The scale bars in the inset images indicate 20 µm. The data are presented as the means ± SDs. * p <0.05, ** p <0.01 compared with the respective control mice, as determined by Fisher's exact probability test or Student's t test.

Article Snippet: Additionally, ATN‐161 (1 mg/kg, MCE, HY‐13535A) was employed to disrupt the tumor–bone matrix interaction mediated by ITGA5.

Techniques: Staining, Immunofluorescence, Inhibition, Derivative Assay, Control

Journal: bioRxiv

Article Title: Endothelial MRG15 Is a Mechanosensitive Suppressor of Atherosclerosis

doi: 10.64898/2026.01.28.702256

Figure Lengend Snippet: Endothelial Mrg15 Deletion promotes inflammation in atherosclerosis. (A) UMAP plots showed Single-cell RNA sequencing (scRNA-seq) analyzed in the carotid arteries of mice at 4 weeks after partial carotid ligation (PCL) surgery from the mice in Mrg15 fl/fl mice (n=6) and Mrg15 iECKO mice(n=6), color-coded for cell types. (B) The proportion of cell type in Mrg15 fl/fl mice and Mrg15 iECKO mice. (C) Heatmap of selected tran transcription factor (TF) activities inferred with DoRothEA from gene expression data generated in EC from Mrg15 fl/fl mice and Mrg15 iECKO mice. (D) Gene Ontology analysis of upregulated genes in EC from Mrg15 iECKO mice compared to Mrg15 fl/fl mice. (E) Kyoto Encyclopedia of Genes and Genomes analysis of upregulated genes in EC from Mrg15 iECKO mice compared to Mrg15 fl/fl mice. (F) UMAP plots showed single-cell transcriptomes analyzed in EC from Mrg15 fl/fl mice and Mrg15 iECKO mice, color-coded for cell types, split by sample origins. (G) Left: Trajectory analysis in EC UMAPs demonstrated EC subtype. Middle and right: Trajectory analysis in EC UMAPs demonstrates pseudo-time, split by sample origins. (H) Trajectory analysis in EC UMAPs demonstrated Icam1 expression, split by sample origins. (I) Representative result and quantitative analysis of ICAM1 staining of LCA sections from the mice in Mrg15 fl/fl mice (n=6) and Mrg15 iECKO mice(n=6) (Scale bar, 50 µm). (J) Representative result and quantitative analysis of ICAM1 staining of LCA sections from the mice in Mrg15 WT mice (n=6) and Mrg15 iECOE mice(n=6) (Scale bar, 50 µm). (K) Real time-qPCR analysis of the expression of Mrg15 , Itga5 , Icam1 , Vcam1 and Ccl5 from the endothelium in Mrg15 fl/fl mice (n=6) and Mrg15 iECKO mice (n=6). (L) Real time-qPCR analysis of the expression of Mrg15 , Itga5 , Icam1 , Vcam1 , Ccl5 from the endothelium in Mrg15 WT mice (n=6) and Mrg15 iECOE mice (n=6). The statistical analysis was performed by two-tailed Student t test for J, Mrg15 , Itga5 , Icam1 and Ccl5 in K, and Ccl5 in L, by Welch’s t test for I, Ccl5 in K, and Mrg15 , Itga5 , Icam1 in L, by Mann-Whitney test for Vcam1 in L.

Article Snippet: Membranes were blocked and then incubated overnight at 4°C with primary antibodies against MRG15 (Cell Signaling Technology, #14098), ITGA5 (Proteintech, #10569-1-AP), ICAM1 (Cell Signaling Technology, #4915), β-actin (Proteintech, #66069-1-Ig), and GAPDH (Proteintech, #60004-1-Ig).

Techniques: RNA Sequencing, Ligation, Gene Expression, Generated, Expressing, Staining, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Endothelial MRG15 Is a Mechanosensitive Suppressor of Atherosclerosis

doi: 10.64898/2026.01.28.702256

Figure Lengend Snippet: MRG15 Decreases ICAM and ITGA5 Expression and Monocyte Adhesion to ECs Induced by Disturbed Flow (A) Real time-qPCR analysis of the expression of Mrg15, Itga5, and Icam1 in primary mouse endothelial cells (MECs) of Mrg15 fl/fl mice (n=6) and Mrg15 iECKO mice (n=6). (B) RT-qPCR analysis of the expression of Mrg15, Itga5, and Icam1 in MECs of Mrg15 WT mice (n=6) and Mrg15 iECOE mice (n=6). (C) Western blot analysis of MRG15, ITGA5, ICAM1, and β-ACTIN levels in human umbilical vein endothelial cells (HUVECs) transfected with control shRNA or MRG15 shRNA adenoviruses for 48 hours and then subjected to STA or oscillatory shear stress (OSS) (6 ± 0.5 dyn/cm 2 ; 1 Hz; n=3) for 12 hours (n=3). (D) Real time-qPCR analysis of ITGA5 and ICAM1 levels in HUVECs transfected with control shRNA or MRG15 shRNA adenoviruses for 48 hours and then subjected to STA or OSS (6 ± 0.5 dyn/cm 2 ; 1 Hz; n=3) for 12 hours (n=3). (E) Representative images of monocyte-endothelial adhesion. HUVECs transfected with control shRNA or MRG15 shRNA adenoviruses for 48 hours and then subjected to STA or OSS (6 ± 0.5 dyn/cm 2 ; 1 Hz; n=3) for 12 hours (n=3) (Scale bar, 50 µm). (F) Western blot analysis of MRG15, ITGA5, ICAM1, and β-ACTIN levels in HUVECs transfected with Ad- GFP or Ad- MRG15 for 48 hours and then subjected to STA or OSS (6 ± 0.5 dyn/cm 2 ; 1 Hz; n=3) for 12 hours (n=3). (G) Real time-qPCR analysis of ITGA5 and ICAM1 levels in HUVECs transfected with Ad- GFP or Ad- MRG15 for 48 hours and then subjected to STA or OSS (6 ± 0.5 dyn/cm 2 ; 1 Hz; n=3) for 12 hours (n=3). (H) Representative images of monocyte-endothelial adhesion. HUVECs transfected with Ad- GFP or Ad- MRG15 for 48 hours and then subjected to STA or OSS (6 ± 0.5 dyn/cm 2 ; 1 Hz; n=3) for 12 hours (n=3) (Scale bar, 50 µm). (I) Representative images of monocyte-endothelial adhesion. Endothelial cells were isolated from Mrg15 fl/fl mice and Mrg15 iECKO mice, treated with PBS or human TNF-α (tumor necrosis factor-α; 10 ng/mL) for 24 hours (n=3) (Scale bar, 50 µm). The statistical analysis was performed by two-tailed Student t test for Mrg15 in A, by Welch’s t test for Itga5 and Icam1 in A, and B, by two-way ANOVA followed by Tukey’s post test for C, D, E, F, G, H, and I.

Article Snippet: Membranes were blocked and then incubated overnight at 4°C with primary antibodies against MRG15 (Cell Signaling Technology, #14098), ITGA5 (Proteintech, #10569-1-AP), ICAM1 (Cell Signaling Technology, #4915), β-actin (Proteintech, #66069-1-Ig), and GAPDH (Proteintech, #60004-1-Ig).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Control, shRNA, Shear, Isolation, Two Tailed Test

Journal: bioRxiv

Article Title: Endothelial MRG15 Is a Mechanosensitive Suppressor of Atherosclerosis

doi: 10.64898/2026.01.28.702256

Figure Lengend Snippet: MRG15 recruits PRC2 to repress transcription of ICAM1 and ITGA5 (A) Representative snapshots of Cleavage Under Targets and Tagmentation (CUT&Tag) tracks for MRG15 in human umbilical vein endothelial cells (HUVECs) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) tracks at the promoters of ITGA5 and ICAM1 in HUVECs transfected with control shRNA or MRG15 shRNA adenoviruses for 48 hours. (B) Coimmunoprecipitation of MRG15, EZH2, SUZ12, and EED in HUVECs. (C) left: chromatin immunoprecipitation followed by qPCR (ChIP-qPCR) analysis validated the enrichment of EZH2 at the promoters of ITGA5 and ICAM1 in HUVECs transfected with control shRNA or MRG15 shRNA adenoviruses for 48 hours (n = 4). right: ChIP-qPCR analysis validated the enrichment of H3K27me3 at the promoters of ITGA5 and ICAM1 in HUVECs transfected with control shRNA or MRG15 shRNA adenoviruses for 48 hours (n = 4). (D) Real time-qPCR analysis of ITGA5 and ICAM1 levels. HUVECs were transfected with Ad- GFP or Ad- MRG15 for 48 hours and then exposed to DMSO or EZH2 inhibitor GSK126 (1 μM) for 24 hours (n=3). (E) Western blot analysis of MRG15, ITGA5, ICAM1, and β-ACTIN levels. HUVECs were transfected with Ad- GFP or Ad- MRG15 for 48 hours and then exposed to DMSO or EZH2 inhibitor GSK126 (1 μM) for 24 hours (n=3). The statistical analysis was performed by two-way ANOVA followed by Tukey’s post test for D and E, by Mann-Whitney test for NC in CHIP-EZH2 and ITGA5 in CHIP-H3K27me3 in C, by two-tailed Student t test for ITGA5 in CHIP-EZH2 and NC in CHIP-H3K27me3 in C, by Welch’s t test for ICAM1 in CHIP-EZH2 and ICAM1 in CHIP-H3K27me3 in C .

Article Snippet: Membranes were blocked and then incubated overnight at 4°C with primary antibodies against MRG15 (Cell Signaling Technology, #14098), ITGA5 (Proteintech, #10569-1-AP), ICAM1 (Cell Signaling Technology, #4915), β-actin (Proteintech, #66069-1-Ig), and GAPDH (Proteintech, #60004-1-Ig).

Techniques: Sequencing, Transfection, Control, shRNA, Chromatin Immunoprecipitation, ChIP-qPCR, Western Blot, MANN-WHITNEY, Two Tailed Test