Altered luminal pH of the Golgi apparatus in GPHR deficient primary cultured neurons. Altered luminal pH of the Golgi apparatus in GPHR deficient primary cultured neurons. A, Immunoblot analysis in primary cultured neurons from GPHRF/F mice with adenovirus (AV)-based exogenous expression of GFP or Cre recombinase. Nine days after infection, total cell lysates were subjected to immunoblot analysis for GPHR and actin. Data are representative of three independent experiments. B, Quantitation of mRNA levels of GPHR in primary cultured neurons manipulated, as described in A. Data are presented as the mean ± SEM; n = 6 for each group; **p < 0.01 (unpaired Student’s t test). C, Fluorescence images of pHluorin-Golgi in primary cultured neurons from GPHRF/+ and GPHRF/F mice with AV-based exogenous expression of Cre recombinase. Two days after infection, pHluorin-Golgi-IRES-mKate2-Golgi was transfected. Fluorescence images were taken seven days after transfection. Data are representative of three independent experiments. Scale bar: 20 µm. D, Quantification of the pHluorin/mKate2 fluorescence ratio in primary cultured neurons manipulated as described in C. Data are presented as the mean ± SEM; n = 5 for each group; **p < 0.01 (unpaired Student’s t test). Yu-shin Sou et al. eNeuro 2019;6:ENEURO.0427-18.2019 ©2019 by Society for Neuroscience