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. 2020 Jan 7;11:19. doi: 10.1038/s41467-019-13842-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Time-lapse imaging demonstrating the spread of calcium transients among cells (left), average calcium transient curve (ΔF/F0) of all cells with calcium transients in one microscope field (right). Initiating cell (white arrows), direction of propagation (yellow arrows). Synchronicity of calcium transients is highlighted by red triangle. Scale Bar: 10 µm. b Left: network of synchronized calcium transients in the indicated wild-type and BCL9 knockout cells. Each dot represents one cell, the colored dots indicate the number of calcium wave events during 30 mins. Edged-linked dots represent synchronized calcium transients, colored edge indicates the timing of synchronized calcium transients. Right: synchronicity of global calcium transients in the indicated cells was calculated in wild-type and BCL9 knockout by FluoroSNNAP, n.d. non-detectable. P values were calculated using Student’s t test. ****P < 0.0001. c Frequency of calcium waves occurring in the indicated wild-type and BCL9 knockout RKO cells in the absence or presence of Verapamil or EDTA. P values were calculated using Student’s t test. *P < 0.05; **P < 0.01; ns: not significant. d Peak height of calcium transients in wild-type and BCL9 knockout RKO cells. P values were calculated using Student’s t test *P < 0.05. e Network of calcium transients spreading (left) and ΔF/F0 of synchronized “secondary waves” (right) after wound scratching in wild-type and BCL9 knockout RKO cells. Each circle represents one cell, colors represent times of simultaneous calcium transients. Edged-linked dots represent synchronized calcium transients, colored edge indicates times of synchronized calcium transients. Red triangle: “secondary waves”; Black line: wound edge. Scale bar: 5 µm. f ΔF/F0 and heatmap showing display of “secondary waves” in Colo320 but not in DLD-1 cells. g The frequency spectrum of calcium waves of indicated ROI in parts e and f. h Co-IP of the indicated proteins in RKO cells treated for 2 h with vehicle (−) or verapamil (+). i Time-course IF depicting changes in BCL9 staining area in cells located adjacent or distant from scratched edge in vehicle (left) or Verapamil (right) treated RKO cells. P values were calculated using Student’s t test. ****P < 0.0001; n.s.: not significant. Data are displayed as mean ± SD. Scale bar: 1 µm. See also Supplementary Figs. 11 and 12. Source data are provided as a Source Data file.