Figure 7. Pax3-3′UTR contains conserved miR-1/miR-206–binding sites. (A) Mouse Pax3 gene structure. Numbered boxes denote each exon. White boxes denote the 5′UTR and the shorter 3′UTR. Black boxes denote coding regions. The gray box denotes the longer 3′UTR containing two putative miR-1/miR-206–binding sites (M1 and M2). There are two stop codons and two polyA signal sequences (polyA1 and polyA2) in the mouse Pax3 gene. (B) Sequences of two putative miR-1/miR-206–binding sites (M1 and M2) and their flanking regions. Core sequences for miR-1/miR-206 and consensus sequences are denoted by bold letters. (C) Similar levels of expression of the Pax3 coding region and the longer 3′UTR were detected in MyoD−/− myoblasts by RT-PCR. β-Actin was monitored as a loading control. (D) MyoD-regulated miRNA expression levels were compared between wild-type (wt) and MyoD−/− myoblasts by RT-PCR. miR-24 was monitored as a loading control. (E) miR-1 and -206 expression were compared between wild-type myoblasts infected with a lentivirus vector expressing shRNA for MyoD and control shRNA (Cont) vector or MyoD−/− myoblasts infected with a lentivirus vector expressing MyoD (Ect-MyoD) and a control empty vector by RT-PCR. miR-24 was monitored as a loading control. (F) Pax3, Pax7, Bcl-2, and Bcl-xL gene expression were compared between MyoD−/− myoblasts transfected with pre–miR-1, pre–miR-206, and the control pre-miRNA by RT-PCR. β-Actin was monitored as a loading control. (G) Pax3, Pax7, Bcl-2, and Bcl-xL gene expression were compared between wild-type myoblasts transfected with anti–miR-1 (anti-1), anti–miR-206 (anti-206), anti–miR-1 + anti–miR-206 (anti-1 + 206), and the control RNA by RT-PCR. β-Actin was monitored as a loading control. (D and E) All PCR products are ∼90-bp long.
Figure 7.

Pax3-3′UTR contains conserved miR-1/miR-206–binding sites. (A) Mouse Pax3 gene structure. Numbered boxes denote each exon. White boxes denote the 5′UTR and the shorter 3′UTR. Black boxes denote coding regions. The gray box denotes the longer 3′UTR containing two putative miR-1/miR-206–binding sites (M1 and M2). There are two stop codons and two polyA signal sequences (polyA1 and polyA2) in the mouse Pax3 gene. (B) Sequences of two putative miR-1/miR-206–binding sites (M1 and M2) and their flanking regions. Core sequences for miR-1/miR-206 and consensus sequences are denoted by bold letters. (C) Similar levels of expression of the Pax3 coding region and the longer 3′UTR were detected in MyoD−/− myoblasts by RT-PCR. β-Actin was monitored as a loading control. (D) MyoD-regulated miRNA expression levels were compared between wild-type (wt) and MyoD−/− myoblasts by RT-PCR. miR-24 was monitored as a loading control. (E) miR-1 and -206 expression were compared between wild-type myoblasts infected with a lentivirus vector expressing shRNA for MyoD and control shRNA (Cont) vector or MyoD−/− myoblasts infected with a lentivirus vector expressing MyoD (Ect-MyoD) and a control empty vector by RT-PCR. miR-24 was monitored as a loading control. (F) Pax3, Pax7, Bcl-2, and Bcl-xL gene expression were compared between MyoD−/− myoblasts transfected with pre–miR-1, pre–miR-206, and the control pre-miRNA by RT-PCR. β-Actin was monitored as a loading control. (G) Pax3, Pax7, Bcl-2, and Bcl-xL gene expression were compared between wild-type myoblasts transfected with anti–miR-1 (anti-1), anti–miR-206 (anti-206), anti–miR-1 + anti–miR-206 (anti-1 + 206), and the control RNA by RT-PCR. β-Actin was monitored as a loading control. (D and E) All PCR products are ∼90-bp long.

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