Figures & data
Figure 1. Transfection efficiency assay of miR-23b-3p on pituitary cells of Yanbian yellow cattle. (a) The level of miR-23b-3p in pituitary cells transfected with the mimics (miR-23b-3p group) and mimics control substance (NC group) of miR-23b-3p. Compared with NC group, the column marked by ** showed significant difference (P<0.01); (b) The level of miR-23b-3p in pituitary cells transfected with the inhibitor (miR-23b-3p-in group) and inhibitor control substance (iNC group) of miR-23b-3p. U6 was used as an internal reference.
Figure 2. Effect of miR-23b-3p on GH mRNA transcription level in pituitary cells of Yanbian yellow cattle. (a) The relative transcription level of GH mRNA in pituitary cells transfected with miR-23b-3p mimics. Mimics (miR-23b-3p-mi group) and mimics reference substance (NC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with NC group, the column marking** showed extremely significant difference (P<0.01); (b) The relative transcription level of GH mRNA in pituitary cells transfected with miR-23b-3p inhibitor. Inhibitor (miR-23b-3p-in group) and inhibitor reference substance (iNC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with iNC group, the column marking* showed significant difference (P<0.05). β-actin was used as an internal reference.
Figure 3. Effect of miR-23b-3p on GH protein expression level in pituitary cells of Yanbian yellow cattle. (a) The relative expression level of GH protein in pituitary cells transfected with miR-23b-3p mimics. Mimics (miR-23b-3p-mi group) and mimics control substance (NC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with NC group, the column marking** showed extremely significant difference (P<0.01); (b) The relative expression level of GH protein in pituitary cells transfected with miR-23b-3p inhibitor. Inhibitor (miR-23b-3p-in group) and inhibitor control substance (iNC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with iNC group, the column marking* showed significant difference (P<0.05). β-actin was used as an internal reference. Electrophoresis of the GH and β-actin gene fragment was from two separate gels.
Figure 4. Target relationship between miR-23b-3p and POU1F1.
Figure 5. Luciferase activity determination. (a) The changes of luciferase activity after PirGLO-POU1F1-3′UTR normal plasmid co-transfecting with miR-23b-3p mimics (miR-23b-3p-mi group) and mimics control substance (NC group) for 48 h. Compared with NC group, the column marking** showed extremely significant difference (P < 0.01); (b) The changes of luciferase activity after PirGLO-POU1F1-3′UTR mutant plasmid co-transfecting with miR-23b-3p mimics (miR-23b-3p-mi group) and mimics control substance (NC group) for 48 h. Compared with NC group, the column without marking**or* showed no significant difference (P >0.05).
Figure 6. Effect of miR-23b-3p on transcription level of POU1F1 mRNA in pituitary cells of Yanbian yellow cattle. (a) The relative transcription level of POU1F1 mRNA in pituitary cells transfected with miR-23b-3p mimics. Mimics (miR-23b-3p-mi group) and mimics control substance (NC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with NC group, the column marking** showed extremely significant difference (P<0.01); (b) The relative transcription level of POU1F1 mRNA in pituitary cells transfected with miR-23b-3p inhibitor. Inhibitor (miR-23b-3p-in group) and inhibitor control substance (iNC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with iNC group, the column marking** showed extremely significant difference (P<0.01). β-actin was used as an internal reference.
Figure 7. Effect of miR-23b-3p on POU1F1 protein expression level in pituitary cells of Yanbian yellow cattle. (a) The relative expression level of POU1F1 protein in pituitary cells transfected with miR-23b-3p mimics. Mimics (miR-23b-3p-mi group) and mimics control substance (NC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with NC group, the column marking** showed extremely significant difference (P<0.01); (b) The relative expression level of POU1F1 protein in pituitary cells transfected with miR-23b-3p inhibitor. Inhibitor (miR-23b-3p-in group) and inhibitor control substance (iNC group) of miR-23b-3p were transfected into pituitary cells of Yanbian yellow cattle, with three replicates in each group. Compared with iNC group, the column marking** showed significant difference (P<0.01). β-actin was used as an internal control. Electrophoresis of the POU1F1 and β-actin gene fragment was from two separate gels.
