Fig 3. Representative TIRFM images of hRPTCs growing on GEM™ 3D microcarriers.

NBCe2 is labeled with a red Alexa 568 fluorescent dye. WT hRPTCs were incubated in vehicle (VEH) (A) and imaged at 70 nm into the plasma membrane to determine intramembranous presence of NBCe2. After 24 h of exposure to high intracellular sodium there was no significant increase in membranous localization of NBCe2 as determined by counting punctate fluorescent spots (B) (quantified in Fig 4). In HV hRPTCs NBCe2 is also expressed in the membrane in VEH treated hRPTCs to a similar level as WT VEH (C) but is increased when the HV hRPTCs were exposed to high intracellular sodium (D)(quantified in Fig 4).