Figure 4.

Y14 deficiency compromises megakaryocyte-maturation-related gene expression and impairs platelet function

(A) The diagram shows the procedure of RNA-seq analysis in control or Y14-depleted HEL cells that were treated or untreated with PMA (see panel B).

(B) HEL cells were transduced with lentivirus expressing shY14 (+) or control shRNA (−) for 48 h at a multiplicity of infection of 5, followed by mock-treated or PMA (5 nM) treatment for 3 days. RT-qPCR analysis of the indicated genes in HEL cells that were transfected with shRNA and then mock-treated or treated with PMA as in panel (A). The experiments were performed in triplicate (mean ± SEM; ∗p < 0.05, ∗∗p < 0.01).

(C) Whole blood of Con and Rbm8aKO^MK mice was collected and stained for IgG, GP-VI, GP-IX, α2bβ3, and CD41 (Con: blue; Rbm8aKO^MK: red). Platelets were gated by forward scattering/side scattering characteristics. Representative flow cytometry profiles showing the indicated glycoproteins or integrins.

(D) Blood was mock-treated (resting; dashed lines) or treated with PAR4 agonist (solid lines) and subsequently stained with an antibody against JON/A (phycoerythrin), fibrinogen, vWF, or P-selectin. Flow cytometry profiles of platelet glycoproteins are shown as in panel (C).