Fig. 5

Identification of PLCβ1 as a direct target of ssc-mir-423-5p in ST cells. a Binding sites for ssc-mir-423-5p in the 3ˈ-UTR of PLCβ1 predicted by TargetScan. Bold font indicate sequences that were mutated to abolish the interaction between ssc-mir-423-5p and PLCβ1 3’UTR. b Luciferase activity was analyzed after co-transfecting pmirGLO-PLCβ1–3′-UTR or pmirGLO-PLCβ1–3′-UTR -mut and ssc-mir-423-5p mimic or mimic NC into ST cells at 24 h. c Luciferase activity was analyzed after co-transfecting pmirGLO-PLCβ1–3′-UTR or pmirGLO-PLCβ1–3′-UTR -mut and ssc-mir-423-5p inhibitor or inhibitor NC into ST cells at 24 h. d PLCβ1 mRNA levels were detected at 48 h after swine cells were transfected with ssc-mir-423-5p mimic or mimic NC. e PLCβ1 mRNA levels were detected at 48 h after swine cells were transfected with ssc-mir-423-5p inhibitor or inhibitor NC. f Western blotting analysis was used to detect PLCβ1 protein expression levels at 72 h after ST cells were transfected with ssc-mir-423-5p mimic and mimic NC. g Western blotting analysis was used to detect PLCβ1 protein expression levels at 72 h after ST cells were transfected with ssc-mir-423-5p inhibitor and inhibitor NC. h The quantification of PLCβ1 protein. *P < 0.05, **P < 0.01, ***P < 0.001, PLCβ1, phospholipase C beta 1; UTR, Untranslated Region; ST, swine testis; NC, negative control; N.S., nonsignificant