Fig. 1

Behavior of ALDH7A1-depleted cells. a Immunoblot of BJ-4F3 cells transduced to express two independent shRNAs targeting ALDH7A1 mRNA (sh-1 and sh-2). Control 1 (C-1) indicates cells transduced with the empty vector. Control 2 (C-2) expressed a non-targeting shRNA. The blot was probed with anti-ALDH7A1. Anti-ACTIN was used as loading control. b Proliferation of BJ-4F3 cells treated as in (a). Cell number was measured by counting DAPI labeled nuclei. X-axis: time in hours, y-axis: relative cell number. Data represent average ± standard error of the mean (SEM) from 3 independent experiments normalized against number of plated cells. ns: the difference was not statistically significant. The two-tailed Mann Whitney test was used to calculate p-values. c Scratch assays to measure cell migration. Images show cells at t = 0 and 24 h. The empty area is shaded red for better visibility. Scale bar: 1000 μm. Average migration distance is shown in μm after 24 h for three independent experiments ± SEM in the panel below. The two-tailed Mann Whitney test was used to calculate p-values. d Matrigel transwell migration assay to measure cell invasiveness. Images show cells labeled with DAPI. Left panels show all cells on the top and bottom surface of the assay well after 24 h. Right panels show cells that successfully migrated through the matrix, which was removed for imaging. The percent of cells that crossed the gel barrier is shown below (average of 3 independent experiments ± SEM). The two-tailed Mann Whitney test was used to calculate p-values