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. 2019 Jul 5;132(13):jcs230052. doi: 10.1242/jcs.230052

Fig. 1.

Fig. 1.

zDHHC15 is expressed during early stages of neocortical development and in Golgi compartments of cultured excitatory and inhibitory neurons. (A,B) Western blot and quantification of zDHHC15 protein levels in the hippocampus (Hc), cerebral cortex (C), hindbrain (Hb) or cerebellum (Cb) of mouse brain at E17, P10 and P90. Levels of zDHHC15 decreased from E17 to P90 in these regions of mouse brain. n=4 tissue samples per time point, *P<0.05, **P<0.001, one-way ANOVA with Tukey's post-hoc test, means±s.e.m. (C) Representative confocal images of 13 DIV rat hippocampal cultures. All neurons, identified by using anti-MAP2 antibody (white arrows), expressed zDHHC15 (yellow arrows) – including inhibitory GAD65-positive neurons (white arrowheads). GFAP-positive glial cells (green arrows) exhibited weak to no zDHHC15 immunostaining. White arrows denote DAPI-stained nuclei that are positive for zDHHC15, whereas green arrows denote DAPI-stained nuclei that are negative for zDHHC15. (D) Western blot of lysates from biotinylated neurons at 13 DIV probed for zDHHC5, zDHHC8 and zDHHC15; the latter is not localized at the membrane. n=3 cultures. (E) zDHHC15 colocalizes with the cis-medial Golgi marker giantin. (F-H) Microimages (F) showing 13 DIV rat hippocampal neurons stained for zDHHC15 and the synaptic markers PSD-95 and gephyrin. zDHHC15 colocalizes with a small subset of PSD-95 or gephyrin puncta. zDHHC15 and PSD-95 images were masked as per Fig. S2E. n=10 neurons, three cultures. Scale bars: 100 µm (C), 20 µm (E, top) and 10 μm (E, bottom; F).