Fig. 4
ALDH7A1 targets the transport function of BARS. Source data are provided as a Source Data file. a HeLa cells were subjected to subcellular fractionation to obtain total membrane vs. cytosol fractions. Both fractions were then subjected to co-immunoprecipitation experiments to detect the association of ALDH7A1 with BARS; n = 2 independent experiments with a representative result shown. b Confocal microscopy was performed to detect the colocalization of ALDH7A1 with different organelle markers; scale bar, 10 µm (2 µm in inset); n = 3 independent experiments with representative images shown. c Quantitation of the colocalization results above; n = 3 independent experiments with ten fields of cells examined in each experiment. d HeLa cells were transfected with GFP-tagged NES-BARS followed by confocal imaging across the nuclear portion of the cell; scale bar, 10 µm; n = 2 independent experiments with a representative image shown. e HeLa cells were fractionated into nuclear vs. cytoplasmic fractions, followed by immunoblotting for proteins as indicated. Lamin B serves as a marker of the nucleus, while actin serves as a marker of the cytoplasm; n = 2 independent experiments with a representative result shown. f HeLa cells were treated as indicated, and then the mRNA level of E-cadherin, which has been a model transcript for studies on transcription repression by BARS/CtBP, is quantified; n = 3 independent experiments. g HeLa cells were treated as indicated, and then colocalization of VSVG-KDELR with a cis-Golgi marker (giantin) coupled with kinetic analysis was performed. Mean with standard deviation is shown; *p < 0.05, paired two-tailed Student’s t-test; n = 3 independent experiments. h HeLa cells were treated as indicated, and then colocalization of dextran with an early endosome marker (EEA1) coupled with kinetic analysis was performed. Mean with standard deviation is shown; *p < 0.05, paired two-tailed Student’s t-test; n = 3 independent experiments
