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. Author manuscript; available in PMC: 2018 Nov 15.
Published in final edited form as: Int J Cancer. 2017 Aug 8;141(10):2062–2075. doi: 10.1002/ijc.30909

Figure 3. miR-130b activates CDC42 through direct target, ARHGAP,1 in Ewing sarcoma.

Figure 3

(A) G-LISA assay of TC71 cells overexpressing miR-130b. Values represent the average of triplicate samples. P-values were determined comparing scramble control lysates to miR-130b mimic lysates. *P<0.01, CDC42 western blot of TC71 scrambled control and miR-130b mimic transfected cells. (B) Representation of the seed pairing between miR-130b and Arhgap1 3′UTR. Bottom represents mutated nucleotides for luciferase assay. (C) qPCR and western blot of ARHGAP1 expression in TC71 cells ectopically overexpressing miR-130b mimic. P-values were determined comparing scramble control to miR-130b overexpression using Student’s t-test *P<0.05 (D) qPCR and western blot of ARHGAP1 expression in MHH-ES1 cells expressing anti-mir to miR-130b. P-values were determined comparing scramble control to anti-miR-130b using Student’s t-test *P<0.05 (E) Luciferase assay of TC71 cells co-transfected with either miR-130b mimic or scrambled control and wildtype 3′UTR ARHGAP1 plasmid or seed mutant plasmid. P-values were determined comparing scramble control to miR-130b overexpression using Student’s t-test *P<0.05. (F) Kaplan-Meier scan for ARHGAP1 derived from clinically annotated human Ewing Sarcoma gene expression database (dataset Mixed Ewing Sarcoma - Savola - 117 - MAS5.0 - u133p2 “http://hgserver1.amc.nl/cgi-bin/r2/main.cgi”. High expression (blue line) and low expression (red line) are compared. Scr = TC71 scrambled control ES cells, 130b = TC71 miR-130b mimic ES cells, anti-130b= anti-miR-130b oligo ES cells.