miR-125a-5p directly regulates L-plastin expression. (A) Real-time quantitative polymerase chain reaction (qPCR) quantification of MK LCP1 mRNA after LNA inhibition of miR-125a-5p (125a LNA) shown as fold change vs negative LNA control (negC) (n = 5). (B) Representative immunoblot of MK L-plastin after miR-125a-5p inhibition. (C) Fold changes of densitometric quantification of L-plastin immunoblots after miR-125a-5p inhibition vs negC, normalized to actin (n = 4). (D) Real-time qPCR quantification of MK LCP1 mRNA fold change after miR-125a-5p (125a Lenti) or control (conL) lentiviral overexpression (n = 8). (E) Representative immunoblot of MK L-plastin after miR-125a-5p overexpression. (F) Fold changes of densitometric quantification of L-plastin immunoblots after miR-125a-5p vs conL overexpression, normalized to actin (n = 7). (G) Luciferase reporter constructs with WT or mutated (Mut) 3′ UTR of LCP1 (left) were cotransfected into HEK293T with a pre-miR precursor of miR-125a-5p (mimic) or a nontargeting miR negative control (ntc) and assayed for luciferase activity (n = 5). (H-I) In vivo murine Lcp1 mRNA levels were assessed by real-time qPCR from mice injected with LNAs as in Figure 3. Fold changes are shown for miR-125a-5p vs negC LNA groups. (H) Purified mouse platelets (n = 7). (I) BM from mouse femurs (n = 8). AU, arbitrary unit.