Figure 1.
FN inhibits FCL formation in a local manner. (A) U2Os-AP2-GFP cells plated on non-coated or FN-coated dishes were imaged using total internal reflection fluorescence (TIRF) microscopy at 1-s intervals for 5 min. Images represent 15-s time projections, and kymographs represent the 5-min time-lapse videos. (B) FCL proportions of U2Os-AP2-GFP cells plated on PLL-, BSA-, FN-, VTN-, Col I-, Col IV-, and LN111-coated or non-coated dishes and imaged using TIRF microscopy, at 1-s intervals for 5 min. N (videos): non-coated (nc) = 32, PLL 3μg/ml = 6, BSA 20 μg/ml = 15, FN 10 µg/ml = 18, FN 20 µg/ml = 14, VTN 20 µg/ml = 14, Col I 20 µg/ml = 14, Col IV 20 µg/ml = 14, LN111 10 µg/ml = 22. Videos were collected from four independent experiments, except for PLL, from two experiments. One-way ANOVA with Tukey’s multiple comparison test. F(8, 183) = 19.11, P < 0.0001. (C) U2Os-AP2-GFP cells plated on FN, VTN, Col I, Col IV, LN111 (all 10 µg/ml) -coated or non-coated dishes and stained for FN. Representative TIRF images. (D) Quantification of FN fluorescent intensity from samples in C. N (images): FN = 15 images, VTN/Col I/LN111/non-coated = 21 images; Col IV = 17 images. Results were obtained from one representative experiment. Similar results were observed in four individual experiments. F(5, 110) = 338.5, P < 0.0001. (E) U2Os-AP2-GFP cells were plated on FN/glass patterned imaging dishes and imaged with TIRF microscopy at 1-s intervals for 5 min. Representative images are 15-s time projections. (F) Representative 5-min kymograph of time-lapse videos in E. (G) Quantification of FCL proportions in E. n = 23 videos, collected from four individual experiments. Two-tailed Student’s t test, P < 0.001. (H) A schematic illustration of the results shown in this figure. Data are the mean ± SD, ns. non-significant P value; * P value < 0.05; P value < 0.01; *** P value < 0.001. Scale bars, 10 µm.

FN inhibits FCL formation in a local manner. (A) U2Os-AP2-GFP cells plated on non-coated or FN-coated dishes were imaged using total internal reflection fluorescence (TIRF) microscopy at 1-s intervals for 5 min. Images represent 15-s time projections, and kymographs represent the 5-min time-lapse videos. (B) FCL proportions of U2Os-AP2-GFP cells plated on PLL-, BSA-, FN-, VTN-, Col I-, Col IV-, and LN111-coated or non-coated dishes and imaged using TIRF microscopy, at 1-s intervals for 5 min. N (videos): non-coated (nc) = 32, PLL 3μg/ml = 6, BSA 20 μg/ml = 15, FN 10 µg/ml = 18, FN 20 µg/ml = 14, VTN 20 µg/ml = 14, Col I 20 µg/ml = 14, Col IV 20 µg/ml = 14, LN111 10 µg/ml = 22. Videos were collected from four independent experiments, except for PLL, from two experiments. One-way ANOVA with Tukey’s multiple comparison test. F(8, 183) = 19.11, P < 0.0001. (C) U2Os-AP2-GFP cells plated on FN, VTN, Col I, Col IV, LN111 (all 10 µg/ml) -coated or non-coated dishes and stained for FN. Representative TIRF images. (D) Quantification of FN fluorescent intensity from samples in C. N (images): FN = 15 images, VTN/Col I/LN111/non-coated = 21 images; Col IV = 17 images. Results were obtained from one representative experiment. Similar results were observed in four individual experiments. F(5, 110) = 338.5, P < 0.0001. (E) U2Os-AP2-GFP cells were plated on FN/glass patterned imaging dishes and imaged with TIRF microscopy at 1-s intervals for 5 min. Representative images are 15-s time projections. (F) Representative 5-min kymograph of time-lapse videos in E. (G) Quantification of FCL proportions in E. n = 23 videos, collected from four individual experiments. Two-tailed Student’s t test, P < 0.001. (H) A schematic illustration of the results shown in this figure. Data are the mean ± SD, ns. non-significant P value; * P value < 0.05; P value < 0.01; *** P value < 0.001. Scale bars, 10 µm.

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