Fig. 1

Generation of transgenic mice expressing V5-tagged PACER in neurons. A Graphical scheme of microinjection strategy. B Genotyping of PACER transgenic mice. PCR detection of HB9-PACER-V5 transgene (single band, 334 bp) in heterozygous PACER-Tg mice. Wild-type mice do not present amplified products for these primers. C, D mRNA levels were determined by qPCR in PACER-Tg (n = 7) versus non-Tg mice (n = 7). C Primers detecting mouse Pacer and human PACER with equal specificity were used to detect total Pacer/PACER mRNA levels in cortex, hippocampus, spinal cord, muscle, or liver. Fold change of mRNA levels was calculated using β-Actin mRNA levels as a reference. D Primers detecting specifically mouse Pacer mRNA or human PACER mRNA or both with equal specificity were used to determine expression levels of endogenous versus exogenous PACER mRNA. β-Actin mRNA levels were determined as a reference. E Detection of PACER in motor neurons of the lumbar spinal cord of PACER-Tg mice by immunofluorescence and confocal microscopy. F Western blot detection of autophagy proteins Beclin1, p62 and LC3-II in the lumbar spinal cord of PACER-Tg (n = 5) versus non-Tg (n = 5) mice. β-Actin protein levels were used as a loading control. G Quantification of protein levels of Beclin1, p62 and LC3-II relative to β-Actin detected in F.  In E and F statistical analysis using Mann–Whitney U-test was performed. Data is presented as means ± S.E.M. p value: p > 0.05: n.s., non-significant, *p < 0.05; **p < 0.01