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. 2015 Jun;100(6):757–767. doi: 10.3324/haematol.2014.118497

Figure 5.

Figure 5.

CD9 mediates PMF megakaryocyte migration in response to CXCL12. (A) Expression of CXCR4 on CD41+ PMF megakaryocytes (Mk) derived from PMF circulating CD34+ cells (D8–10 of culture) by flow cytometry and microscopy as compared to Mk derived from healthy donors (unmobilized peripheral blood or bone marrow). (B) CD9 and CXCR4 coexpression on CD41low and CD41high PMF cells. (C) Effect of surface CD9 ligation by anti-CD9 monoclonal antiboby (mAb) (clone-SYB; 10 μg/mL) on CXCR4 and CXCL12 transcript expression normalized to RPL38 by QRT-PCR in Mk derived from PMF circulating CD34+ cells (D8–10 of culture). (D) Effect of silencing CD9 using siRNA CD9 (1 μg/106 cells) in Mk derived from PMF circulating CD34+ cells (D8–10 of culture) on the CD9 transcript and protein expression levels analyzed by QRT-PCR (at 48 h) and by flow cytometry (at 72 h), respectively, and on the percentage of migrating PMF Mk in a Boyden chamber in response to CXCL12 (100 ng/mL) as compared to passive diffusion and on the phospho-FAK (Tyr 576/577) expression level (MFI).