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. 2020 Nov 11;11:529757. doi: 10.3389/fgene.2020.529757

FIGURE 6.

FIGURE 6

miR-149-5p suppresses goat hair follicle stem cell apoptosis. (A) mRNA expression of antiapoptotic gene (Bcl2) at 24 h, 48 h, 72 h, and 96 h after transfection with NC, miR-149-5p mimics, anti-NC, and miR-149-5p inhibitors in GM as determined by RT-qPCR. (B,C) mRNA expression of apoptosis marker genes (Caspase3 and Caspase9) at 24 h, 48 h, 72 h, and 96 h after transfection with NC, miR-149-5p mimics, anti-NC, and miR-149-5p inhibitors in GM as determined by RT-qPCR. (D) Protein expression of Bcl2 (1:1000 dilution), Caspase3 (1:1000 dilution), and Caspase9 (1:1000 dilution) was measured at 24 h, 48 h, 72 h, and 96 h after transfection with NC, miR-149-5p mimics, anti-NC, and miR-149-5p inhibitors in GM. β-actin (1:500 dilution) was used as an internal control. (E) Relative Bcl2 protein expression with NC, miR-149-5p mimics, anti-NC, and miR-149-5p inhibitors in GM at 24 h, 48 h, 72 h, and 96 h. (F) Relative Caspase3 protein expression with NC, miR-149-5p mimics, anti-NC, and miR-149-5p inhibitors in GM at 24 h, 48 h, 72 h, and 96 h. (G) Relative Caspase9 protein expression with NC, miR-149-5p mimics, anti-NC, and miR-149-5p inhibitors in GM at 24 h, 48 h, 72 h, and 96 h. (H,I) Hair follicle stem cells were transfected with NC or miR-149-5p mimics in GM, and cell apoptosis was analyzed 48 h after transfection by Annexin V-FITC/PI binding followed by flow cytometry. (J,K) Hair follicle stem cells were transfected with anti-NC and miR-149-5p inhibitors in GM, and cell apoptosis was analyzed 48 h after transfection by Annexin V-FITC/PI binding followed by flow cytometry. The results from each group are shown as the mean ± SEM of three independent replicates. Independent-samples t-tests were used for statistical analysis. Asterisks indicate significant differences. No asterisk means P > 0.05, *P < 0.05, and **P < 0.01.