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Osteosarcoma cell-derived exosomal-miR-487a promotes macrophages polarization toward to M2-like phenotype. ( A ) After preprocessingGSE33382, only samples of osteosarcoma were retained. Analysis of osteosarcoma tissues using CIBERSORT, followed by determination of the percentage of macrophages in tumor tissues. ( B ) Correlation between miR-487a expression and distinct gene signatures of various immune cells. ( C ) FCM was used to identify the macrophage marker CD14. ( D ) qRT-PCR was used to assess the the expression of M2 macrophages markers. ( E ) FCM was used to measure the expression of the M2 macrophage marker <t>(CD163)</t> following treatment with MG-63 exosomes and HOS exosomes. ( F ) qRT-PCR and ( G ) ELISA were used to detect the expression of M2 markers (Arg-1, Ym1 and <t>CD163)</t> and M1 markers (CD86 and iNOS) following treatment with MG-63 exosomes and HOS exosomes. ( H , I ) The expression of M2 markers (Arg-1, Ym1 and CD163) and M1 markers (CD86 and iNOS) were detected after treated with MG-63 anti-miR-487a exosomes and HOS anti-miR-487a exosomes with qRT-PCR ( H ) and ELISA ( I ) (* p <0.05; ** p <0.01; *** p <0.001)
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Osteosarcoma cell-derived exosomal-miR-487a promotes macrophages polarization toward to M2-like phenotype. ( A ) After preprocessingGSE33382, only samples of osteosarcoma were retained. Analysis of osteosarcoma tissues using CIBERSORT, followed by determination of the percentage of macrophages in tumor tissues. ( B ) Correlation between miR-487a expression and distinct gene signatures of various immune cells. ( C ) FCM was used to identify the macrophage marker CD14. ( D ) qRT-PCR was used to assess the the expression of M2 macrophages markers. ( E ) FCM was used to measure the expression of the M2 macrophage marker <t>(CD163)</t> following treatment with MG-63 exosomes and HOS exosomes. ( F ) qRT-PCR and ( G ) ELISA were used to detect the expression of M2 markers (Arg-1, Ym1 and <t>CD163)</t> and M1 markers (CD86 and iNOS) following treatment with MG-63 exosomes and HOS exosomes. ( H , I ) The expression of M2 markers (Arg-1, Ym1 and CD163) and M1 markers (CD86 and iNOS) were detected after treated with MG-63 anti-miR-487a exosomes and HOS anti-miR-487a exosomes with qRT-PCR ( H ) and ELISA ( I ) (* p <0.05; ** p <0.01; *** p <0.001)
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Osteosarcoma cell-derived exosomal-miR-487a promotes macrophages polarization toward to M2-like phenotype. ( A ) After preprocessingGSE33382, only samples of osteosarcoma were retained. Analysis of osteosarcoma tissues using CIBERSORT, followed by determination of the percentage of macrophages in tumor tissues. ( B ) Correlation between miR-487a expression and distinct gene signatures of various immune cells. ( C ) FCM was used to identify the macrophage marker CD14. ( D ) qRT-PCR was used to assess the the expression of M2 macrophages markers. ( E ) FCM was used to measure the expression of the M2 macrophage marker (CD163) following treatment with MG-63 exosomes and HOS exosomes. ( F ) qRT-PCR and ( G ) ELISA were used to detect the expression of M2 markers (Arg-1, Ym1 and CD163) and M1 markers (CD86 and iNOS) following treatment with MG-63 exosomes and HOS exosomes. ( H , I ) The expression of M2 markers (Arg-1, Ym1 and CD163) and M1 markers (CD86 and iNOS) were detected after treated with MG-63 anti-miR-487a exosomes and HOS anti-miR-487a exosomes with qRT-PCR ( H ) and ELISA ( I ) (* p <0.05; ** p <0.01; *** p <0.001)

Journal: Cancer Cell International

Article Title: The sEVs miR-487a/Notch2/GATA3 axis promotes osteosarcoma lung metastasis by inducing macrophage polarization toward the M2-subtype

doi: 10.1186/s12935-024-03488-x

Figure Lengend Snippet: Osteosarcoma cell-derived exosomal-miR-487a promotes macrophages polarization toward to M2-like phenotype. ( A ) After preprocessingGSE33382, only samples of osteosarcoma were retained. Analysis of osteosarcoma tissues using CIBERSORT, followed by determination of the percentage of macrophages in tumor tissues. ( B ) Correlation between miR-487a expression and distinct gene signatures of various immune cells. ( C ) FCM was used to identify the macrophage marker CD14. ( D ) qRT-PCR was used to assess the the expression of M2 macrophages markers. ( E ) FCM was used to measure the expression of the M2 macrophage marker (CD163) following treatment with MG-63 exosomes and HOS exosomes. ( F ) qRT-PCR and ( G ) ELISA were used to detect the expression of M2 markers (Arg-1, Ym1 and CD163) and M1 markers (CD86 and iNOS) following treatment with MG-63 exosomes and HOS exosomes. ( H , I ) The expression of M2 markers (Arg-1, Ym1 and CD163) and M1 markers (CD86 and iNOS) were detected after treated with MG-63 anti-miR-487a exosomes and HOS anti-miR-487a exosomes with qRT-PCR ( H ) and ELISA ( I ) (* p <0.05; ** p <0.01; *** p <0.001)

Article Snippet: To investigate the impact of sEVs derived from MG-63 and HOS cells on macrophage expression of iNOS, CD86, and CD163.ELISA was performed using a human Arg-1 (#EK1316, Multi Sciences, Hangzhou, China), Ym1 (#DY2599, R&D system, USA), CD163 (#DC1630, R&D system), iNOS (#CB11694-Hu, Coibo Bio, Shanghai, China) and CD86 (#CB15455-Hu, Coibo Bio, Shanghai, China) ELISA kit as protocol.

Techniques: Derivative Assay, Expressing, Marker, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Exosomal-miR-487a targets Notch2 and induces M2 polarization by activating GATA3. ( A ) TargetScan is used for miR-487a target gene prediction. ( B ) Macrophages’ relative Notch2 mRNA expression. ( C ) Western blot was used to confirm the expression of Notch2 in macrophages was discovered following treatment with miR-487a mimics or inhibitor. ( D ) Expression of Notch2 in macrophages after treatment with MG-63 exosomes and HOS exosomes. ( E ) Expression of Notch2, GATA3, and IL-4 proteins in macrophages following treatment with MG-63 exosomes and HOS exosomes. ( F-G ) Macrophages were transfected either with miR-487a plus si-Notch2 or miR-487a mimics. The expression of M2 markers (Arg-1, Ym1, and CD163) was assessed using qRT-PCR ( F ) and FCM ( G ). ( H ) Western blot analysis was used to identify components of the Notch2/GATA3 pathway. ( I , J ) MG-63 exosomes, MG-63 exosomes plus si-Notch2, HOS exosomes, or HOS exosomes plus si-Notch2 were applied to macrophages. ( K ) FCM was used to confirm the expression of CD163, an M2 marker (* p <0.05; ** p <0.01; *** p <0.001)

Journal: Cancer Cell International

Article Title: The sEVs miR-487a/Notch2/GATA3 axis promotes osteosarcoma lung metastasis by inducing macrophage polarization toward the M2-subtype

doi: 10.1186/s12935-024-03488-x

Figure Lengend Snippet: Exosomal-miR-487a targets Notch2 and induces M2 polarization by activating GATA3. ( A ) TargetScan is used for miR-487a target gene prediction. ( B ) Macrophages’ relative Notch2 mRNA expression. ( C ) Western blot was used to confirm the expression of Notch2 in macrophages was discovered following treatment with miR-487a mimics or inhibitor. ( D ) Expression of Notch2 in macrophages after treatment with MG-63 exosomes and HOS exosomes. ( E ) Expression of Notch2, GATA3, and IL-4 proteins in macrophages following treatment with MG-63 exosomes and HOS exosomes. ( F-G ) Macrophages were transfected either with miR-487a plus si-Notch2 or miR-487a mimics. The expression of M2 markers (Arg-1, Ym1, and CD163) was assessed using qRT-PCR ( F ) and FCM ( G ). ( H ) Western blot analysis was used to identify components of the Notch2/GATA3 pathway. ( I , J ) MG-63 exosomes, MG-63 exosomes plus si-Notch2, HOS exosomes, or HOS exosomes plus si-Notch2 were applied to macrophages. ( K ) FCM was used to confirm the expression of CD163, an M2 marker (* p <0.05; ** p <0.01; *** p <0.001)

Article Snippet: To investigate the impact of sEVs derived from MG-63 and HOS cells on macrophage expression of iNOS, CD86, and CD163.ELISA was performed using a human Arg-1 (#EK1316, Multi Sciences, Hangzhou, China), Ym1 (#DY2599, R&D system, USA), CD163 (#DC1630, R&D system), iNOS (#CB11694-Hu, Coibo Bio, Shanghai, China) and CD86 (#CB15455-Hu, Coibo Bio, Shanghai, China) ELISA kit as protocol.

Techniques: Expressing, Western Blot, Transfection, Quantitative RT-PCR, Marker

Biomarker concentrations

Journal: Journal of the Endocrine Society

Article Title: Transgender Women With Suppressed Testosterone Display Lower Burden of Coronary Disease Than Matched Cisgender Men

doi: 10.1210/jendso/bvae120

Figure Lengend Snippet: Biomarker concentrations

Article Snippet: Soluble CD14 (sCD14, R and D Systems catalog No. DC140, RRID:AB_3095885), soluble CD163 (sCD163, R and D Systems catalog No. DC1630, RRID:AB_3096052), interleukin-6 (IL-6, R and D Systems catalog No. HS600C, RRID:AB_2893335), fatty acid-binding protein-4 (FABP-4, (ImmunoDiagnostics catalog No. 31030, RRID:AB_2933962), high-molecular-weight adiponectin (R and D Systems catalog No. DRP300, RRID:AB_2783020), insulin (R and D Systems catalog No. DINS00, RRID:AB_3073852), endothelin-1 (ET-1, R and D Systems catalog No. DET100, RRID:AB_3099470), and vascular cell adhesion molecule-1 (VCAM-1, R and D Systems catalog No. DVC00, RRID:AB_2941366) were analyzed by Quantikine ELISA (enzyme-linked immunosorbent assay), and soluble tumor necrosis factor receptor type (sTNFR, R and D Systems catalog No. LUCAM726, RRID:AB_3099473) I/II, interleukin-8 (IL-8, R and D Systems catalog No. DY208, RRID:AB_2892143), and plasminogen activator inhibitor-1 (PAI-1, R and D Systems catalog No. LOBM1786, RRID:AB_3099472) were measured by Luminex assay.

Techniques: Biomarker Assay