Fig 2: CircCOL1A1 regulates miR-149-5p and CMTM3 expression in goat hair follicle stem cells. (A,B) Expression of miR-149-5p in hair follicle stem cells after cotransfection with circCOL1A1-si and miR-149-5p-in (A) and with circCOL1A1 and miR-149-5p (B). (C,D) The relative expression of CMTM3 in hair follicle stem cells after cotransfection with circCOL1A1-si and miR-149-5p-in (C) and with circCOL1A1 and miR-149-5p (D). (E,G) CMTM3 protein expression in hair follicle stem cells after cotransfection with circCOL1A1-si and miR-149-5p-in. (F,H) CMTM3 protein expression in hair follicle stem cells after cotransfection with circCOL1A1 and miR-149-5p. (I) Fold enrichment of CMTM3 and AR determined by RT-q-PCR following the RIP assay with Ago-2 antibody and IgG antibody (negative control).

Fig 3: CircCOL1A1 acts as a sponge for miR-149-5p in goat hair follicle stem cells. (A) The predicted circCOL1A1 binding site sequence on miR-149-5p using RNAhybrid (v2.1.2) software. (B) The predicted secondary structure and base pair probabilities for circCOL1A1 and miR-149-5p using RegRNA v2.0 software. (C) miR-149-5p mimics were co-transfected with the circCOL1A1-wild-type or circCOL1A1-mut vector into HEK293T cells. (D) The pcDNA3.1 (+)-miR-149-5p plasmid was co-transfected with the circCOL1A1-wild-type or circCOL1A1-mut vector into HEK293T cells. (E) The miR-149-5p Inhibitors were co-transfected with the circCOL1A1-wild-type or circCOL1A1-mut vector into HEK293T cells. (F) A schematic diagram of the miR-149-5p biosensor structure. (G) The miR-149-5p biosensor was co-transfected with miR-149-5p mimics or circCOL1A1-overexpressing plasmid into 293T cells. (H) Fold enrichment of circCOL1A1 and miR-149-5p determined by RT-q-PCR following the RIP assay with Ago-2 and IgG antibodies (negative control). (I) The expression of miR-149-5p after circCOL1A1 overexpression and inhibition using the RT-q-PCR assay. (J) Expression abundance of circCOL1A1, miR-149-5p, and CMTM3 in cervical spine skin tissues from NQ and SQ goats.