Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: MM cell-derived ETV1 fosters M2 polarization of TAMs. ( A ) THP-1 cells were incubated with 350 nM PMA for 24 h to induce M0 macrophage differentiation. The morphology of M0 macrophages was photographed with a microscope. ( B ) The proportion of CD68 + cells in M0 macrophages were detected by flow cytometry. ( C ) M2 polarization of TAMs was measured by Transwell co-culture system. M0 macrophages were seeded into the lower chamber, and MM cells were seeded into the upper chamber. ( D ) The levels of CCL2 in MM cells was tested by qPCR and ELISA. ( E ) After 48 h of co-culture, the expression of CD163, Arg-1 and CD206 in M0 macrophages was determined by qPCR. ( F ) The proportion of CD206 + cells and MFI in M0 macrophages were examined by flow cytometry. Data are mean ± SD. #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: Derivative Assay, Incubation, Microscopy, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: ETV1 facilitates tumor growth and M2 polarization of macrophages in vivo. ( A ) The development of tumors was determined by the in vivo fluorescence imaging system. ( B ) H&E staining was used to detect tumor formation in bone tissues of mice. (Scale bar = 100 μm). ( C ) Immunohistochemical staining was used to examine the expression of human ETV1 in bone marrow tissues. (Scale bar = 50 μm). ( D ) Flow cytometry was used to test the proportion of mouse-derived M2 phenotypic macrophages (F4/80 + CD206+/F4/80+) in M0 macrophages (F4/80+) in bone marrow tissues. ( E ) The expression of human CCL2 in bone marrow tissues was detected by western blot. Data are mean ± SD. ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: In Vivo, Fluorescence, Imaging, Staining, Immunohistochemical staining, Expressing, Flow Cytometry, Derivative Assay, Western Blot

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: RBMS1 knockdown counteracts the effect of ETV1 overexpression on MM cell proliferation and M2 polarization of TAMs. ( A ) siRNAs targeting RBMS1 were transfected into RPMI8226 cells. After 48 h, the knockdown efficiency of RBMS1 in cells was verified by qPCR. ( B ) Cell viability was measured by CCK-8 assay. ( C ) Cell cycle distribution was detected by flow cytometry. ( D ) The expression of cyclinD1 in cells was detected by western blot. ( E ) The levels of CCL2 in the cell supernatant were determined by ELISA. ( F ) M2 polarization of TAMs was tested by Transwell co-culture system. M0 macrophages were seeded into the lower chamber, and MM cells were seeded into the upper chamber. After 48 h of co-culture, the expression of CD163 and CD206 in M0 macrophages were determined by qPCR. ( G ) The proportion of CD206 + cells and MFI in M0 macrophages were examined by flow cytometry. Data are mean ± SD. #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: Knockdown, Over Expression, Transfection, CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Co-Culture Assay