Fig. 2

Combined treatment with Fe-EO and Thy-EO (Mix-EO) suppresses MMP9 expression and melanoma cell migration. (A and B) Murine B53, MEF, B16F1, B16F10, MS-5 (A) and human HEK293 HS-5, A431, SKMEL28, and HUVEC cells (B) were analyzed for MMP9 mRNA expression by qPCR (n = 3/cell line). Expression was compared to the expression in B53 (mouse) or HEK293 (human) cells. C Human MMP9 expression levels in tumor (T) and adjacent normal tissues (N) derived from SKCM patients. Data were retrieved from the TCGA database and analyzed by Timer. D, E Fold change in MMP9 expression in B16F10 (D) and A431 (E) cells treated with indicated Mix-EO concentrations as determined by qPCR. Expression was compared to untreated controls. D, E Fold change in MMP9 expression in B16F10 (D) and A431 € cells treated with indicated concentrations of Mix-EO for 24 h as determined by qPCR. Expression was compared to the expression in control (DMSO)-treated cells. F Fold change in P65 expression in B16F10 and A431 cells treated with indicated concentrations of Mix-EO for 24 h (n = 6/group). G Representative immunoblots for p65, mouse MMP9, and b-actin (loading controls) using B16F10 cell lysates of indicated treatment groups. H–J Microscopic images 24 h after wound healing closure in B16F10 cultures treated with DMSO (control) or indicated EOs at indicated concentrations (n = 3/goup). I Quantification of B16F10 cells that migrated in the scraped area indicated by thick lines per high power fields (HPF). A total of 5 × HPFs were counted. Data are expressed as mean ± SEM (unpaired Student's t test or one-way ANOVA *p < .05)