Fig. 9
GlCHC and GlACLC foci associate to Giardia PV/PECs with different stoichiometries. A An epitope-tagged GlCHC reporter was used to localize the foci of GlCHC deposition at the cell periphery beneath the PM, by STED microscopy. B These foci are well-resolved with STED microscopy (left graph). C The same was done with an epitope-tagged GlACLC reporter, and D the foci resolved as in B (right graph). E The foci were segmented with ilastik. Areas were determined and diameters calculated in an automatic procedure assuming spherical geometry. GlCHC foci present an average diameter of 134 ± 36.6 nm (N = 4524) with a 95% confidence interval between 132 and 135 nm. GlACLC foci average a diameter of 159 ± 48.8 nm (N = 984) with a 95% confidence interval between 156 and 162 nm. GlACLC foci are larger than GlCHC in significant statistical manner (p-value < 0.0001, t-Student test). F GlCHC foci associate in different stoichiometry to different classes of PV/PECs. Spherical PV/PECs are either not associated to clathrin or with just one focus: a mean of 0.488 ± 0.159 foci per spherical with a 95% confidence interval between 0.403 and 0.573 foci per spherical organelle. Tubular PV/PECs associate with one focus of GlCHC: a mean of 1.15 ± 0.287 with a 95% confidence interval between 0.994 and 1.3 foci per tubular organelle. Polymorphic PV/PECs associate with 3 or more GlCHC foci with an average of 3.85 ± 1.14 with a 95% confidence interval between 3.25 and 4.46 foci per polymorphic organelle. These distributions are statistically significant with a p-value < 0.0001 (ANOVA analysis)