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Fig. 9 | Molecular Neurodegeneration

Fig. 9

From: A Christianson syndrome-linked deletion mutation (∆287ES288) in SLC9A6 disrupts recycling endosomal function and elicits neurodegeneration and cell death

Fig. 9

Over-acidification of endosomes containing NHE6 ΔES. AP-1 cells were transiently transfected with 3FNHE6HA WT and ΔES and endosomal delivery was assessed 36 h post-transfection. Anti-Flag M2 primary and FITC-conjugated Fab secondary antibodies were bound to live cells for 1 h on ice. The temperature was raised to 37 °C for 30 or 60 min and endosomal pH was measured by fluorescence ratio imaging (FRIA). a In situ calibration of FITC fluorescence as a function of vesicular pH was performed by clamping the vesicular pH between 4 and 7.5 as described in “Methods”. b Measurement of vesicular pH as a function of time upon internalization of 3FNHE6HA WT and ΔES. The pH values represent the mean ± S.E.M. and the number of vesicles (~250 to 1000 vesicles) analyzed from a representative experiment are shown. c Graphical plot of the median vesicular pH as a function of time derived from four experiments (mean ± S.E.M.)

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Molecular Neurodegeneration

ISSN: 1750-1326