Fig 1: PLAU increases colony formation of SW480 cells. The colonies were fixed with methanol for 30 min and stained with 0.1% crystal violet for 20 min. The formed colonies of SW480 cells (magnification ×1) increased after in vitro overexpression of PLAU. The representative images of formed colonies in the NC RNA of PLAU-inhibitor (A), PLAU-inhibitor (B), the NC for PLAU-mimic (C), and PLAU-mimic group (D). (E) The number of formed colonies compared by bar chart. **, P<0.01; compared with the NC RNA of PLAU-inhibitor; ##, P<0.01, compared with the NC of PLAU-mimic. PLAU, plasminogen activator, urokinase; NC, negative control.
Fig 2: PLAU-mimic increases the invasion ability of CRC cells. (A) The images of invaded cells in the NC RNA of PLAU-inhibitor (PLAU-NC), PLAU-inhibitor, the NC for PLAU-mimic (PLAU-NC), and PLAU-mimic group. Cells were fixed in 4% paraformaldehyde for 20 minutes and stained with 0.1% crystal violet for 15 minutes. (B) The number of invaded cells compared by bar chart. **, P<0.01; compared with the NC RNA of PLAU-inhibitor; ##, P<0.01, compared with the NC of PLAU-mimic. CRC, colorectal cancer; PLAU, plasminogen activator, urokinase; NC, negative control.
Fig 3: The DEGs analysis and bioinformatics analysis of the GSE156355 and GSE184093 dataset. (A) The volcano plot of DEGs in GSE156355 dataset. (B) The heatmap of DEGs in GSE156355 dataset. (C,D) The volcano plot and heatmap of the differentially expressed mRNAs in GSE184093 dataset. (E,F) The identified pathways GO analysis at the BP level in GSE156355 and GSE184093 dataset. (G,H) The identified downregulated pathways. (I) The KEGG enrichment analysis of DEGs. (J,K) The GSEA analysis of GSE156355 and GSE184093 dataset. (L) The expression of PLAU analyzed using the GEPIA database, PLAU was highly expressed in CRC. (M,N) The DFS and OS curves. *, P<0.05. DEGs, differentially expressed genes; GO, Gene Ontology; BP, biological processes; FC, fold change; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, gene set enrichment analysis; GEPIA, Gene Expression Profiling Interactive Analysis; PLAU, plasminogen activator, urokinase; TPM, Transcripts Per Million mapped reads; CRC, colorectal cancer; DFS, disease-free survival; OS, overall survival.
Fig 4: The migration ability CRC cells measured using wound healing after PLAU was increased. Cells were fixed in 4% paraformaldehyde for 20 minutes and stained with 0.1% crystal violet for 20 minutes. PLAU-mimic enhances the migration of CRC cells. (A) From left to right, the representative images of scratch in cells transfected the NC RNA of PLAU-inhibitor (PLAU-NC), PLAU-inhibitor, the negative control for PLAU-mimic (PLAU-NC), and PLAU-mimic in 0, 24, and 48 hours. Scale bar =200 µm. (B) Column chart comparing different scratch distances in the four groups. *, P<0.05; **, P<0.01, compared with the NC RNA of PLAU-inhibitor; #, P<0.05; ##, P<0.01, compared with the NC of PLAU-mimic. CRC, colorectal cancer; PLAU, plasminogen activator, urokinase; NC, negative control.
Fig 5: The protein expressions of p-Src, p-ERK1/2, MMP-2, MMP-3, MMP-9 CyclinD1, and CyclinA2 in SW480 cells measured by western blot. (A) Representative western images of above proteins in cells transfected with the NC RNA of PLAU-inhibitor (PLAU-NC), PLAU-inhibitor, the NC for PLAU-mimic (PLAU-NC), and PLAU-mimic. (B) The relative expression of p-Src, p-ERK1/2, MMP-2, MMP-3, MMP-9, CyclinD1, CyclinA2, and GADPH in each group. **, P<0.01. ns, not significant. PLAU, plasminogen activator, urokinase; NC, negative control; MMP, matrix metallopeptidase.
Supplier Page from Abcam for Recombinant Human PLAU Protein (Active)