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. 2012 Apr 6;287(22):17975–17984. doi: 10.1074/jbc.M111.310458

FIGURE 3.

FIGURE 3.

Human keratinocytes lacking α6β4 integrin expression display adhesion and migration defects. In A, control keratinocytes (HEK) and α6 shRNA clone 1 cells were assayed for adhesion to laminin-332- or collagen I-coated dishes at 1 h after plating. The number of adherent cells was measured by reading the absorbance at 570 nm. The average absorbance for HEK was taken as 100%. The individual bars on the graph represent the means ± S.E. (n = 3) with samples measured in triplicate for each experiment. p values were derived by comparing samples to HEK (Student's t test). *, p ≤ 0.05. B, vector diagrams depict the individual migration patterns of wild-type keratinocytes (HEK) (n = 11), α6 shRNA clone 1 cells (n = 12), or α6 shRNA clone 1 cells induced to re-express GFP-tagged α6 integrin (+refα6) (n = 12). The cells were plated on glass and tracked over a 2-h period. C, Rac1 activity in control HEK and α6 shRNA clone 1 cells was measured by the G-LISA Rac1 activation assay. The individual bars of the graph represent the means ± S.E. (n = 3) (in each experiment, duplicate samples were assayed). p value was derived by comparing to HEK (Student's t test). *, p ≤ 0.05. D, graph representing the average velocity of cells plated on glass over a 2-h period. The individual bars of the graph represent the means ± S.E. (n ≥ 50 cells). **, p ≤ 0.001. E, vector diagrams depicting the migration pattern of HEK (n = 28), α6 shRNA clone 1 cells (n = 37), or α6 shRNA clone 1 cells infected with retrovirus encoding GFP-tagged α3 integrin (+α3) plated on matrix deposited by HEK (n = 19). The cells were allowed to adhere to the matrix for 2 h, and the migration of cells was subsequently tracked for 2 h. F, graph represents the average migration index (net displacement/total distance) of cells over a 2-h period. The individual bars of the graph represent the means ± S.E. (n ≥ 50 cells). G, graph represents the average velocity of cells plated on pre-formed matrix over a 2-h period. The individual bars of the graph represent the means ± S.E. (n ≥ 50 cells). p values were derived by Student's t test. *, p ≤ 0.05.