Fig 2: ?-Catenin is a novel regulator of HAI-1/SPINT-1. A, gene network analysis based on previously known direct (solid lines) and indirect (dashed lines) biological connections for the identified transcripts with a positive (red) and negative (green) correlation (r-value >0.5) to ?-catenin expression in two, independent lung cancer cell line datasets. Genes with an asterisk were represented more than once in the 152 transcripts correlated to ?-catenin gene expression. B and C, H157 (B) and H1299 (C) cells were transfected with either control or ?-catenin plasmids as described under “Experimental Procedures.” Total RNAs isolated from the transfected cells were employed in quantitative PCR (qPCR) analysis. **, p < 0.01; versus control. D and E, H157 (D) and H1299 (E) cells were transfected with either control or ?-catenin plasmids as described under “Experimental Procedures.” The cell lysates were later probed for the expression of ?-catenin and HAI-1 via immunoblotting with anti-?-catenin and anti-HAI-1 antibodies. F, total cell lysates of A549 cells stably expressing ?-catenin were probed for the expression of ?-catenin, p53, and HAI-1 via immunoblotting with anti-?-catenin, anti-p53, and anti-HAI-1 antibodies. G, H157 cells were co-transfected with ?-catenin plasmids and p53 siRNAs as described under “Experimental Procedures.” The cell lysates were later probed for the expression of ?-catenin, p53, and HAI-1 via immunoblotting with specific antibodies. H, H157 cells were co-transfected with ?-catenin plasmids and HAI-1 siRNAs as described under “Experimental Procedures,” and their cell proliferation rates were determined with an MTS assay. Data represent mean ± S.E. of three independent experiments. #, p < 0.05; versus ?-catenin + Ctrl siRNA control.

Fig 3: HAI-1 is a novel regulator of NSCLC cell proliferation and migration. A and B, HAI-1 was transiently expressed in NSCLC cells (i.e. H157 and H1299 cells). The cell lysates were later probed to detect the expression of HAI-1 by immunobloting with anti-HAI-1 antibodies. C–F, cell proliferation rates of H157 (C and E) and H1299 cells (D and F) transfected with either control or HAI-1 plasmids were determined with either MTS assays (C and D) or clonogenic assays (E and F) as described under “Experimental Procedures.” Data represent mean ± S.E. of three independent experiments. ##, p < 0.01; #, p < 0.05; versus control. G and H, H157 (G) and H1299 (H) cells were transfected with HAI-1 plasmids and allowed to grow to confluence. A 3-mm scrape wound was created in confluent cultures, and cell migration was recorded at 0 and 24 h as described under “Experimental Procedures.” I, H1299 cells were co-transfected with ?-catenin and WT p53 plasmids. Cell migration was later assayed in trans-well inserts as described under “Experimental Procedures.” Top panel represents the number of cells migrated, while representative images were displayed in the bottom panel. ##, p < 0.01; versus control. J, H1299 cells were co-transfected with WT p53 plasmids and HAI-1 siRNAs. Cell migration was later assayed in trans-well inserts as described under “Experimental Procedures.” Top panel represents the number of cells migrated, while representative images were displayed in the bottom panel. ##, p < 0.01; versus control, **, p < 0.01; versus p53 + control siRNA control.