Research Paper Volume 15, Issue 23 pp 13980—13997

Figure 4. Prediction and validation of miR-520d-5p as one of target miRNAs of GPRC5D-AS1. (A) qRT-PCR analyzed gene expression level of GPRC5D-AS1, miR-520d-5p, miR-153-3p and miR-524-5p. Different concentrations of Dex (5 mM, 10 mM and 15 mM) were added in HSMM and incubated for 48 h. Control: HSMM; T-1: 5 mM Dex; T-2:10 mM Dex; T-3:15 mM Dex. ^*P < 0.05, ^**P < 0.01 compared with control group. (B) NC mimic and miR-520d-5p mimic were co-transfected with plasmid psiCHECK2-GPRC5D-AS1-WT luciferase vector or psiCHECK2-GPRC5D-AS1-MUT vector into in human skeletal muscle myoblasts, and the normalized relative luciferase activities (Renilla/firefly) were analyzed. ^*P < 0.05. (C) qRT-PCR analyzed gene expression of miR-520d-5p. HSMM was control group. Dex (15 mM) was added in HSMM to establish atrophy cell model (model group). Empty plasmid (NC group) and GPRC5D-AS1-OE plasmid (lncRNA-OE group) were transfected into atrophy cell model and incubated for 48 h. ^*P < 0.05, ^**P < 0.01 compared with control group; ^#P < 0.05, ^##P < 0.01 compared with model group; ^&P < 0.05, ^&&P < 0.01 compared with NC group.