Fig. 3 | Nature Communications

Fig. 3

From: miR-143/145 differentially regulate hematopoietic stem and progenitor activity through suppression of canonical TGFβ signaling

Fig. 3

Loss of miR-143 and miR-145 activates the TGFβ/Dab2/Smad3 signaling axis. a Measurement of luciferase activity after transfection of 3TP-LUX, a luciferase reporter containing TGFβ-responsive elements, into 293T cells constitutively expressing DAB2 (DAB2-CE) and stimulated with vehicle (Veh) or 5 ng/ml TGFβ. Data are expressed as arbitrary units (AUs, mean ± SEM, n = 3). b Phosphorylation of Smad2/3 (pSmad2/3) in lineage-negative marrow cells of wild-type (WT) and miR-143/145−/− mice stimulated with vehicle or 5 ng/ml TGFβ by intracellular flow cytometry. Data are expressed as geometric mean fluorescence intensity (gMFI, mean ± SEM, n = 3). c Western blot of Dab2 in WT and miR-143/145−/− Lin− marrow isolated by immunomagnetic negative selection. Densitometry data were normalized to Gapdh and are presented as a ratio relative to vector (mean ± SEM, n = 7, P = 0.001)). d Luciferase activity after co-transfection with empty vector or miR-145 overexpression construct with the WT or seed site-mutated 3′-UTR of DAB2 inserted downstream of a luciferase reporter (mean ± SEM, n = 3)

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