Research Paper Volume 13, Issue 2 pp 2700—2726

Figure 5. Confirmation of the quaternary ceRNA network regulating TYR. (A) After transfection with siRNAs of ENST00000606533 in MCs for 48 h, the transfection efficiency of si-ENST00000606533 mix (mixing the three siRNAs) was most significant, and the expression of circ_0091223, miR-1291, and TYR were detected by transfection with si-ENST00000606533 mix. (B) After transfection with siRNAs of circ_0091223 for 48 h, the transfection efficiency of si-circ_0091223 2 was most significant, and the expression of ENST00000606533, miR-1291, and TYR were detected by transfection with si-circ_0091223 2. (C) After transfection with miR-1291 mimics for 48 h, the transfection efficiency, the expression of ENST00000606533, circ_0091223, and TYR were detected. (D) After treated with α-MSH, the MCs were transfected with si-ENST00000606533 mix, or si-circ_0091223 2, or miR-1291 mimics, and the expression of ENST00000606533, circ_0091223, miR-1291, and TYR were detected. (E) After co-transfection with siRNAs of ENST00000606533 and miRNA_1291 inhibitor for 48 h, the expression of ENST00000606533, circ_0091223, miR-1291, and TYR were detected. (F) After co-transfection with siRNA of circ_0091223 and miR-1291 inhibitor for 48 h, the expression of ENST00000606533, circ_0091223, miR-1291, and TYR were detected. (G) After treated with α-MSH and/or si-ENST00000606533 mix, or si-circ_0091223 2, or miR-1291 mimics, the protein level of TYR were detected. (H) After treated with α-MSH and/or si-ENST00000606533 mix, or si-circ_0091223 2, or miR-1291 mimics, the Masson-Fontana melanin staining were detected.