Figure 6. Integrin β1 blocking... | Rockefeller University Press

Figure 6.

$Integrin β1 blocking stimulates FCL and RA formation. (A) U2Os-AP2-GFP cells plated on FN were treated with integrin β1 blocking antibody mab13 (0.3 µg/ml) for 15 and 45 min (or vehicle for 45 min = control) and stained for integrin αvβ5 and p-Pax. Representative TIRF images. (B) Analysis of AP2 signal colocalizing with no markers (non), p-Pax, or integrin αvβ5 over time. N (images): control = 18, mab13 15 min = 13, mab13 45 min = 15, from one representative experiment. Similar results were observed in four independent experiments. One-way ANOVA with Tukey’s multiple comparison. F(2, 119) = 46.68, P < 0.0001. (C) Analysis of fraction of integrin αvβ5 colocalizing with AP2 over time from samples in A. N (images): control = 15, mab13 15 min = 11, mab13 45 min = 12, from one representative experiment. Similar results were observed in four independent experiments. One-way ANOVA with Tukey’s multiple comparison. F(2, 35) = 45.81, P < 0.0001. (D) U2Os-AP2-GFP cells plated on FN were treated with mab13 (0.3 µg/ml) and imaged using TIRF microscopy. 5-min time-lapses with 1-s intervals starting at 0 min (no mab13) and every 5 min after mab13 addition, until 35 min, were acquired. N (videos): 0 min = 11, 5 min = 11, 10 min = 9, 15 min = 8, 20 min = 8, 25 min = 6, 30 min = 7, 35 min = 6. Videos were acquired from two independent experiments. Similar results were observed in five individual experiments. One-way ANOVA with Tukey’s multiple comparison. F(7, 29) = 8.893, P < 0.0001. (E) A schematic illustration of results shown in this figure. Data are the mean ± SD, ns. non-significant P value; * P value<0.05, *** P value < 0.001. Scale bars, 10 µm; insets, 5 µm.$

Integrin β1 blocking stimulates FCL and RA formation. (A) U2Os-AP2-GFP cells plated on FN were treated with integrin β1 blocking antibody mab13 (0.3 µg/ml) for 15 and 45 min (or vehicle for 45 min = control) and stained for integrin αvβ5 and p-Pax. Representative TIRF images. (B) Analysis of AP2 signal colocalizing with no markers (non), p-Pax, or integrin αvβ5 over time. N (images): control = 18, mab13 15 min = 13, mab13 45 min = 15, from one representative experiment. Similar results were observed in four independent experiments. One-way ANOVA with Tukey’s multiple comparison. F(2, 119) = 46.68, P < 0.0001. (C) Analysis of fraction of integrin αvβ5 colocalizing with AP2 over time from samples in A. N (images): control = 15, mab13 15 min = 11, mab13 45 min = 12, from one representative experiment. Similar results were observed in four independent experiments. One-way ANOVA with Tukey’s multiple comparison. F(2, 35) = 45.81, P < 0.0001. (D) U2Os-AP2-GFP cells plated on FN were treated with mab13 (0.3 µg/ml) and imaged using TIRF microscopy. 5-min time-lapses with 1-s intervals starting at 0 min (no mab13) and every 5 min after mab13 addition, until 35 min, were acquired. N (videos): 0 min = 11, 5 min = 11, 10 min = 9, 15 min = 8, 20 min = 8, 25 min = 6, 30 min = 7, 35 min = 6. Videos were acquired from two independent experiments. Similar results were observed in five individual experiments. One-way ANOVA with Tukey’s multiple comparison. F(7, 29) = 8.893, P < 0.0001. (E) A schematic illustration of results shown in this figure. Data are the mean ± SD, ns. non-significant P value; * P value<0.05, *** P value < 0.001. Scale bars, 10 µm; insets, 5 µm.