Fig 1: SERP1 Was the Target Gene of miR-526b-5p in OS Cells(A) The intersection of the differentially expressed mRNAs (log2FC = 1.5 with p = 0.05) of the GEO: GSE37552 dataset and the predicted targets of miR-526b-5p by the TargetScan Human 7.2 database. (B) TargetScan Human 7.2 was used to predict the binding site of the SERP1 3' UTR for miR-526b-5p. (C) The dual-luciferase reporter assay was used to prove the binding site between the SERP1 3' UTR and miR-526b-5p. SERP1-WT, the plasmids contained the binding site of SERP1 3' UTR for miR-526b-5p; SERP1-MUT, the plasmids did not have any biding site of the SERP1 3' UTR for miR-526b-5p. *p < 0.05, **p < 0.001 versus co-transfection of SERP1-WT and NC plasmids. (D) The expression of SERP1 mRNA was detected in normal and tumor tissues. (E) The negative correlation between the expression of SERP1 and miR-526b-5p in 16 OS tissues is shown. (F) Expression of SERP1 in HOS and Saos2 cells after stable transfection of Con, NC, sh-SERP1, miR-526b-5p inhibitor, or sh-SERP1+miR-526b-5p inhibitor. Con, the HOS and Saos2 cells were cultured without any treatments; NC, the HOS and Saos2 cells were transfected with negative control vectors; sh-SERP1, the HOS and Saos2 cells were transfected with sh-SERP1; inhibitor, the HOS and Saos2 cells were transfected with miR-526b-5p inhibitor; sh-SERP1+inhibitor, the HOS and Saos2 cells were co-transfected with sh-SERP1 and miR-526b-5p inhibitor. *p < 0.05, **p < 0.001. (G) The protein level of SERP1 was assessed via western blot assay in HOS and Saos2 cells after stable transfection of Con, NC, sh-SERP1, miR-526b-5p inhibitor, or sh-SERP1+miR-526b-5p inhibitor. Three independent repeated experiments were implemented in all of the assays.
Fig 2: Inhibition of SERP1 Reversed miR-526b-5p Inhibitor-Induced Cell Proliferation, Migration, and Invasion in OS Cells(A) The proliferation of the transfected HOS and Saos2 cells was evaluated by a CCK-8 assay. Con, the HOS and Saos2 cells were cultured without any treatments. NC, the HOS and Saos2 cells were transfected with negative control vectors. Sh-SERP1, the HOS and Saos2 cells were transfected with sh-SERP1. Inhibitor, the HOS and Saos2 cells were transfected with miR-526b-5p inhibitor. Sh-SERP1+inhibitor, the HOS and Saos2 cells were co-transfected with sh-SERP1 and miR-526b-5p inhibitor. *p < 0.05, **p < 0.001 versus control group. #p < 0.05, ##p < 0.001 versus inhibitor group. (B) The proliferation of HOS and Saos2 cells was assessed by colony formation assay. Con, the HOS and Saos2 cells were cultured without any treatments. NC, the HOS and Saos2 cells were transfected with negative control vectors. Sh-SERP1, the HOS and Saos2 cells were transfected with sh-SERP1. Inhibitor, the HOS and Saos2 cells were transfected with miR-526b-5p inhibitor. Sh-SERP1+inhibitor, the HOS and Saos2 cells were co-transfected with sh-SERP1 and miR-526b-5p inhibitor. *p < 0.05, **p < 0.001 versus control group; #p < 0.05, ##p < 0.001 versus inhibitor group. (C) Wound-healing assay was performed to assess the migration ability of the transfected HOS and Saos2 cells. Con, the HOS and Saos2 cells were cultured without any treatments; NC, the HOS and Saos2 cells were transfected with negative control vectors; sh-SERP1, the HOS and Saos2 cells were transfected with sh-SERP1; inhibitor, the HOS and Saos2 cells were transfected with miR-526b-5p inhibitor; sh-SERP1+inhibitor, the HOS and Saos2 cells were co-transfected with sh-SERP1 and miR-526b-5p inhibitor. *p < 0.05, **p < 0.001 versus control group; #p < 0.05, ##p < 0.001 versus inhibitor group. (D) The invasion ability was evaluated by a transwell invasion assay in HOS and Saos2 cells. Con, the HOS and Saos2 cells were cultured without any treatments; NC, the HOS and Saos2 cells were transfected with negative control vectors; sh-SERP1, the HOS and Saos2 cells were transfected with sh-SERP1; inhibitor, the HOS and Saos2 cells were transfected with miR-526b-5p inhibitor; sh-SERP1+inhibitor, the HOS and Saos2 cells were co-transfected with sh-SERP1 and miR-526b-5p inhibitor. *p < 0.05, **p < 0.001 versus control group; #p < 0.05, ##p < 0.001 versus inhibitor group.
Supplier Page from Abcam for Anti-SERP1 antibody