Fig. 6

MiR-144-3p targets and suppresses the expression of PPARGC1B and DUSP16 in the chicken liver. a, b The potential miR-144-3p binding sites in the 3′-UTR of PPARGC1B (a) and DUSP16 (b). The exact location and the sequence of the binding sites are indicated. c–e Luciferase activities driven by the three miR-144-3p binding sites (~ 200 bp) in DF1 cells transfected with the miR-144-3p mimic or mimic NC (negative control). Luciferase activities were measured 48 h after transfection. Error bars represent SEM, n = 3, *p < 0.05, **p < 0.01. f The miR-144 mimic or mimic NC was transfected into chicken primary hepatocytes. After 24 h, qRT-PCR was performed to determine the expression levels of miR-144-3p, PPARGC1B, and DUSP16. g The miR-144-3p inhibitor or inhibitor NC was transfected into chicken primary hepatocytes. After 24 h, qRT-PCR assays were performed to determine the expression levels of miR-144-3p, PPARGC1B, and DUSP16. h, i Expression patterns of miR-144-3p and DUSP16 and PPARGC1B mRNAs in response to 17β-estradiol in the chicken liver according to qRT-PCR analysis. All data are presented as mean ± SEM, n = 6 for the control group and n = 8 for the 17β-estradiol treatment groups. j Comparison between the expression curve of miR-144-3p and the egg production curve of the Lushi green-shelled-egg chickens. Each point in the egg production curve represents the cumulative egg production of 2000 chickens in one week. k, l Expression levels of miR-144-3p and DUSP16 and PPARGC1B mRNAs in the chicken liver at different ages of chickens. Expression levels were determined by qRT-PCR. All data are presented as mean ± SEM