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. 2017 Nov 1;28(22):3013–3028. doi: 10.1091/mbc.E17-01-0059

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© 2017 Chiang et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 6: — Arl4C-FLNa interaction is necessary for cell migration. (A) Representative images of wound-healing and transwell assays of HeLa cells transfected with empty vector, Arl4C, and FLNa siRNA. Confluent monolayers were photographed at the indicated times after wounding. Migration ability was calculated by comparing the area of the wound between 0 and 24 h, as indicated, using Metamorph software. Scale bar = 45 µm. The migrated cells in a field were calculated using ImageJ software after 24 h of migration, as indicated. (B) Immunoblotting analyses were used to verify the level of protein expression in HeLa cells. Total protein (20 µg) was loaded onto a 10-well gel to detect proteins. The percentage of FLNa after siRNA treatment is 23.8 ± 0.67%. (C) Representative images of wound-healing assays of HeLa cells transfected with empty vector, Arl4C, full-length FLNa-WT, -A2, -A10, and FLNa siRNA. Confluent monolayers were photographed at the indicated times after wounding. Migration ability was calculated by comparing the area of the wound between 0 and 16 h, as indicated, using Metamorph software. Scale bar = 45 µm. (D) Immunoblotting analyses were used to verify the level of protein expression in HeLa cells. Total protein (20 µg) was loaded onto a 10-well gel to detect proteins. The percentage of FLNa after siRNA treatment is 37.3 ± 0.6%. (E) Representative images of wound-healing assays of A549 cells transfected with empty vector, full-length FLNa-WT, -A2, -A10, and FLNa siRNA. Confluent monolayers were photographed at the indicated times after wounding. Migration ability was calculated by comparing the area of the wound between 0 and 16 h, as indicated, using Metamorph software. Scale bar = 45 µm. (F) Immunoblotting analyses were used to verify the level of protein expression in A549 cells. Total protein (20 µg) was loaded onto a 10-well gel to detect proteins. The percentage of FLNa after siRNA treatment is 19.7 ± 0.4%. Histograms in A, C, and E: Quantification of migration assays was based on three biological replicates. Scatter plots represent mean ± SD. *, p < 0.05, **, p < 0.005, ***, p < 0.001 (two-tailed Student’s t test).