Figure 2

(A) Loss-of-function (LOF) screens were performed in human primary lung fibroblasts (hLF) or mouse 3T3 fibroblasts by transfecting an antisense RNA against both miR-125a and miR-125b (miR-125a/b-AS), or by microinjecting morpholinos (MO) against pre-mir-125b hairpin precursors (all 3 isoforms) into zebrafish embryos. Gain-of-function (GOF) screens were performed in human SH-SY5Y and mouse N2A neuroblastoma by transfecting the miR-125b duplex into cells in culture, or by coinjecting the miR-125b duplex with the morpholinos against pre-mir-125b into zebrafish embryos. Fold changes in gene expression were measured by qRT-PCR twenty-four hours after transfection or injection, relative to the mock and negative control miRNA or morpholino, and shown as log₂(fold change) using a heat-map. (B) Human: 13 genes were significantly derepressed by a loss of miR-125b, while 20 genes were significantly repressed by a gain of miR-125b, making a total of 22 genes that passed the screen (P<0.05, fold change > 1.3, relative to mock control). (C) Mouse: 11 genes were significantly derepressed by a loss of miR-125b, while 12 genes were significantly repressed by a loss of miR-125b, making a total of 13 genes that passed the screen (P<0.05, fold change > 1.3, relative to mock control). (D) Zebrafish: 13 genes were significantly derepressed by a loss of pre-mir-125b (P<0.05, fold change > 1.3, relative to control MO), while 12 genes were significantly repressed/rescued by a gain of miR-125b (P<0.05, fold change > 1.3, relative to pre-mir-125b MO), making a total of 14 genes that passed the screen. All experiments were performed with at least three biological replicates.