Fig 1: High-grade cancer cell lines are specifically characterized by scattered, peripheral positioning of lysosomes.A Representative images of lysosomes visualized by immunofluorescence staining against the lysosomal-associated membrane protein 1 (LAMP1/CD107a) in normal human urothelium (NHU) and bladder cancer cell lines RT4 (ATCC® HTB-2™), MGHU370, RT11271, KU19-1972, T24, TCCSup73, JMSU174 cells cultured on crossbow-shaped adhesive micropatterns for better comparison. Scale bar is 10 µm. B 3D probabilistic density maps of lysosomes of N cells of NHU, RT4, MGHU3, RT112, KU19-19, T24, TCCSup and JMSU1. The 50% contour visualizes the smallest cellular volume containing 50% of lysosomes. C Nearest neighbor distance (NND) between lysosomes in NHU (n = 76), RT4 (n = 73), MGHU3 (n = 65), RT112 (n = 64), KU19-19 (n = 64), T24 (n = 72), TCCSup (n = 48) and JMSU1 (n = 60). Adjusted p values of testing against NHU condition are RT4: > 0.9999, MGHU3: 0.1943; RT112: < 0.0001; KU19-19: <0.0001; T24: < 0.0001; TCCsup: <0.0001; JMSU1: < 0.0001 in a Kruskal-Wallis test with Dunn’s test for multiple comparisons; ns p > 0.1 and ****p < 0.0001, error bars are SEM. D Average numbers of lysosomes per cell in NHU (n = 76), RT4 (n = 73), MGHU3 (n = 65), RT112 (n = 64), KU19-19 (n = 64), T24 (n = 72), TCCSup (n = 48) and JMSU1 (n = 60). Adjusted p values of testing against NHU condition are RT4: <0.0001; MGHU3: < 0.0001; RT112: > 0.9999; KU19-19: 0.8807; T24: > 0.9999; TCCsup: 0.2068; JMSU1: < 0.0001 in a Kruskal-Wallis test with Dunn’s test for correction for multiple comparisons; ns p > 0.1 and ****p < 0.0001, error bars are SEM. E Average volume of lysosomes in NHU (n = 76), RT4 (n = 73), MGHU3 (n = 65), RT112 (n = 64), KU19-19 (n = 64), T24 (n = 72), TCCSup (n = 48) and JMSU1 (n = 60). Adjusted p values of testing against NHU condition are RT4: 0.1414; MGHU3: < 0.0001; RT112: 0.0048; KU19-19: 0.0110; T24: > 0.9999; TCCsup: 0.0003; JMSU1: < 0.0001 in a Kruskal-Wallis test with Dunn’s test for multiple comparisons; ns p > 0.1, *p < 0.1, ***p < 0.001 and ****p < 0.0001, error bars are SEM.
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