miR-144 selectively inhibits embryonic α-E1 globin expression. (A) Morphology of 22 hpf embryos injected with either miR-144 LNA control (left) or miR-144 LNA oligonucleotides (middle, right). (B) WISH assays for α-E1 globin. A significant increase of α-E1 globin mRNAs was observed in the miR-144 knockdown embryos (middle, arrows), with approximately 8% of miR-144 knockdown embryos showing massive induction (right, arrow). (C,D) WISH assays for alas2 (C) and α-E3 globin (D). A slight increase (alas2 and α-E3) and an ectopic expression (alas2, right panel, arrowhead) were noted. (E-I) WISH assays for β-E1 hemoglobin (E), β-E2 hemoglobin (F), gata-1 (G), scl (H), and l-plastin (I, arrows). No obvious changes were detected. (J) Representative results of genes analyzed by semiquantitative, single-embryo RT-PCR in miR-144 LNA and control-injected embryos (n = 3). (K) Whole-mount immunohistochemistry staining for mitotic phosphorylated histone 3 (pH3). Brown dots indicate cells undergoing mitosis. (L) Apoptotic assay. Arrows indicate red dots and TUNEL-positive cells.