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. 2022 Mar 7;17(1):441–452. doi: 10.1515/med-2022-0441

Figure 2.

Figure 2

miR-195 targeted and decreased the expression of FASN. (a) Target Scan Human 6.0 software predicted that there was a target site of miR-195 on 680–686 bp of 3′-UTR of FASN. (b) Luciferase activity assay showed the following: the luciferase activity in the group psi-FASN Vector + miR-195 was significantly lower than that in the group FASN + negative controls (n = 6; p < 0.001), but there were no significant differences between the group psi-FASN mutVector + miR-195 and the group FASN mut + negative (n = 6: #p = 0.083), and the luciferase activity in the group psi-FASNmut Vector + miR-195 was significantly higher than that in the group psi-FASN Vector + miR-195 (n = 6; **p < 0.001). The error bar represents the standard variation of the luciferase activity in the cells transfected with FASN (or mutant FASN) plasmids and treated with or without miR-195. (c) The expression level of miR-195 was significantly up/downregulated in U2-OS and 143B cells transfected by miR-195/up or miR-195/down lentiviral vector (n = 6). The error bar represents the standard variation of the miR-195 levels in the U2-OS or 143B cells transfected with lentiviruses overexpressing or inhibiting miR-195. (d) The FASN mRNA expression of osteosarcoma cells decreased significantly in the group miR-195/up and significantly upregulated in the group miR-195/down in vitro by RT-PCR analysis (n = 6). The error bar represents the standard variation of the FASN levels in the U2-OS or 143B cells transfected with lentiviruses overexpressing or inhibiting miR-195. (e) The FASN protein expression of osteosarcoma cells decreased significantly in the group miR-195/up and significantly upregulated in the group miR-195/down in vitro by western blot analysis (n = 6).