Fig. 8

HeLa cells overexpressing mutant ADGRL2 present enhanced cell adhesive properties associated to signal transduction alteration. a-c HeLa cells expressing either pcD-Empty (a), CIRL2-Wt (b) or CIRL2-Mt (c) were labelled with the viability marker, cell tracker green (10 μM), and the mortality marker 7-AAD (50 μg/ml) and incubated at room temperature for 90 min in aggregation medium. Note the marked increase of aggregate sizes in CIRL2-Mt expressing cells. d-f HeLa cells expressing either pcD-Empty (d) CIRL2-Wt (e) or CIRL2-Mt (f) were incubated at room temperature for 90 min in aggregation medium containing the PLC inhibitor U73122 (3 μM). Note that inhibition of PLC enhanced homophilic binding of HeLa cells overexpressing CIRL2-Wt. g-i HeLa cells expressing either pcD-Empty (g), CIRL2-Wt (h) or CIRL2-Mt (i) were incubated at room temperature for 90 min in aggregation medium containing α-latrotoxin (1 nM) which prevented cell aggregation. j-l HeLa cells expressing either pcD-GFP-Empty (j), CIRL2-GFP-Wt (k) or CRL2-GFP-Mt (l) were incubated at room temperature for 90 min in aggregation medium. Cells expressing CIRL2 coupled to GFP in C-terminal were not able to aggregate. m Quantification and statistical analysis of the aggregation index. Each value represents the mean (±S.E.M.) of three independent cell-adhesion assays. **, p < 0.01; ***, p < 0.001, ****, p < 0.0001 using one-way ANOVA test