Figure 7.
Figure 7. Functional assays on CHO, MCF-7, HCC1937, and HCC1937 + WT BRCA1 cell lines. (A) Cell spreading: the percentage of spread out cells per field at 3 h after seeding for CHO–wild type (CHO-WT) and CHO stably expressing low (CHO-Low) or high (CHO-High) levels of EGFP-CTD (means ± SD; n = 5). Each pairwise comparison by t test gave P < 0.01. (B) Table comparing mean speeds for different cell populations. CHO-WT and MCF-7 moved slower than their high expressing counterparts CHO-High and MCF-7–High. MCF-7–High moved faster than HCC1937. HCC1937 cells rescued with full-length BRCA1 (HCC1937 + WT BRCA1) displayed a highly significant lower mean speed than their native HCC1937 counterpart. All but one of the mean speed differences were highly significant (P < 0.01). There is no difference in the mean speed of HCC1937 + WT BRCA1 versus MCF-7 (P = 0.1). Means ± SEM for CHO-WT, CHO-Low, CHO-High, HCC1937 + WT, MCF-7, HCC1937, and MCF-7–High are 0.38 ± 0.01, 0.42 ± 0.01, 0.84 ± 0.02, 0.29 ± 0.01, 0.32 ± 0.01, 0.52 ± 0.01, and 0.89 ± 0.02, respectively (n = 4). (C) Motility of CHO cell populations: a speed frequency histogram is shown for CHO-WT, CHO-High, and CHO-Low cell types (means ± SD; n = 4). The CHO-High moved faster than CHO-WT and CHO-Low. CHO-WT and CHO-Low behaved similarly. (D) Motility of MCF-7, HCC1937, HCC1937 + WT BRCA1, and MCF-7–High cell populations: a speed frequency histogram is shown for MCF-7, HCC1937, HCC1937 + WT BRCA1, and MCF-7–High cells (means ± SD; n = 4). HCC1937 and MCF-7–High cells moved faster than HCC1937 + WT BRCA1 and MCF-7. MCF-7–High cells moved faster than HCC1937. Restoring full-length BRCA1 in HCC1937 reversed the hypermotility pattern. X-axis values are upper bounds for each speed bin. (E) Cell tracks for CHO-WT, CHO-High, MCF-7, and MCF-7–High cells are shown, including phase contrast and EGFP-CTD expression. CHO-WT and MCF-7 moved less far than their high expressing counterparts over this 1-h time period imaged at 2-min intervals. (F) Wound healing of CHO-WT, unsorted CHO–EGFP-CTD, and CHO-High cell monolayers. PVIs from seven independent wounds are presented for each cell type. The CHO-High population displayed the highest PVI values, which were correlated with a highly irregular wound margin. CHO-WT and unsorted CHO–EGFP-CTD showed a more regular behavior. Means for CHO-WT, unsorted CHO–EGFP-CTD, and CHO-High were 51, 73, and 209, respectively (horizontal lines). Nonparametric two-tailed Mann–Whitney U tests gave P < 0.05 (CHO-WT vs. unsorted CHO–EGFP-CTD) and P < 0.001 (CHO-WT vs. CHO-High; n = 7).

Functional assays on CHO, MCF-7, HCC1937, and HCC1937 + WT BRCA1 cell lines. (A) Cell spreading: the percentage of spread out cells per field at 3 h after seeding for CHO–wild type (CHO-WT) and CHO stably expressing low (CHO-Low) or high (CHO-High) levels of EGFP-CTD (means ± SD; n = 5). Each pairwise comparison by t test gave P < 0.01. (B) Table comparing mean speeds for different cell populations. CHO-WT and MCF-7 moved slower than their high expressing counterparts CHO-High and MCF-7–High. MCF-7–High moved faster than HCC1937. HCC1937 cells rescued with full-length BRCA1 (HCC1937 + WT BRCA1) displayed a highly significant lower mean speed than their native HCC1937 counterpart. All but one of the mean speed differences were highly significant (P < 0.01). There is no difference in the mean speed of HCC1937 + WT BRCA1 versus MCF-7 (P = 0.1). Means ± SEM for CHO-WT, CHO-Low, CHO-High, HCC1937 + WT, MCF-7, HCC1937, and MCF-7–High are 0.38 ± 0.01, 0.42 ± 0.01, 0.84 ± 0.02, 0.29 ± 0.01, 0.32 ± 0.01, 0.52 ± 0.01, and 0.89 ± 0.02, respectively (n = 4). (C) Motility of CHO cell populations: a speed frequency histogram is shown for CHO-WT, CHO-High, and CHO-Low cell types (means ± SD; n = 4). The CHO-High moved faster than CHO-WT and CHO-Low. CHO-WT and CHO-Low behaved similarly. (D) Motility of MCF-7, HCC1937, HCC1937 + WT BRCA1, and MCF-7–High cell populations: a speed frequency histogram is shown for MCF-7, HCC1937, HCC1937 + WT BRCA1, and MCF-7–High cells (means ± SD; n = 4). HCC1937 and MCF-7–High cells moved faster than HCC1937 + WT BRCA1 and MCF-7. MCF-7–High cells moved faster than HCC1937. Restoring full-length BRCA1 in HCC1937 reversed the hypermotility pattern. X-axis values are upper bounds for each speed bin. (E) Cell tracks for CHO-WT, CHO-High, MCF-7, and MCF-7–High cells are shown, including phase contrast and EGFP-CTD expression. CHO-WT and MCF-7 moved less far than their high expressing counterparts over this 1-h time period imaged at 2-min intervals. (F) Wound healing of CHO-WT, unsorted CHO–EGFP-CTD, and CHO-High cell monolayers. PVIs from seven independent wounds are presented for each cell type. The CHO-High population displayed the highest PVI values, which were correlated with a highly irregular wound margin. CHO-WT and unsorted CHO–EGFP-CTD showed a more regular behavior. Means for CHO-WT, unsorted CHO–EGFP-CTD, and CHO-High were 51, 73, and 209, respectively (horizontal lines). Nonparametric two-tailed Mann–Whitney U tests gave P < 0.05 (CHO-WT vs. unsorted CHO–EGFP-CTD) and P < 0.001 (CHO-WT vs. CHO-High; n = 7).

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