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. 2017 Mar 31;6:e23172. doi: 10.7554/eLife.23172

Figure 6. Ambra1 binding-impaired mutant FAK increases invasion and 3D proliferation, which is rescued by reducing Dynactin 1 or IFITM3 expression levels.

Figure 6.

(A) Invasion assay: SCC FAK-WT, FAK P875A/P881A (AA) and FAK -/- cells were seeded on growth factor-reduced Matrigel in serum-free conditions. After 72 hr, invasion towards a serum gradient was visualised by staining the cells with calcein. n = 8. Error bars, s.e.m. p≤0.01. (B) Representative images of the invasion assay. (C, D) 3D proliferation assay: SCC FAK-WT, FAK P875A/P881A and FAK -/- cells were resuspended in methylcellulose solution in growth medium on a layer of agarose. After nine days, images were taken from 6–10 random fields and colonies were counted. (C) Quantification of the 3D proliferation assay. n = 3. Error bars, s.d. p<0.01. (D) Representative images of the 3D proliferation assay. (E–G) Invasion assay: SCC FAK-WT, FAK P875A/P881A and FAK -/- cells transiently transfected with Dctn1 siRNA were seeded on growth factor-reduced Matrigel in serum-free conditions. After 72 hr, invasion towards a serum gradient was visualised by staining the cells with calcein. (E) Representative images of the invasion assay. (F) Quantification of the invasion assay. n = 6. Error bars, s.e.m. p≤0.01. (G) Quantification of the invasion assay with cells transiently transfected with IFITM3 siRNA. n = 6. Error bars, s.e.m. p≤0.01 (*) and p<0.05 (#).

DOI: http://dx.doi.org/10.7554/eLife.23172.019

Figure 6—source data 1. Mean values of invasion and number of colonies.
Mean percentage and s.e.m. values of the relative invasion of SCC FAK-WT, P875A/P881A and -/- cells (Figure 6A), as well as upon Dctn1 (Figure 6F) and IFITM3 (Figure 6G) knockdown by siRNA are shown. The mean number of colonies and s.d. are shown (Figure 6C).
DOI: http://dx.doi.org/10.7554/eLife.23172.020
elife-23172-fig6-data1.xlsx (43.8KB, xlsx)
DOI: 10.7554/eLife.23172.020