Fig. 1

KLHL17/AF, a brain-specific protein, is colocalized with or adjacent to F-actin in mature neurons. a Functional domains of KLHL17/AF. Both the N- and C-terminal regions of KLHL17/AF have two functions, as indicated. b Expression of KLHL17/AF in diverse adult mouse tissues (3-months-old). A total of 20 μg of tissue lysates from each sample was analyzed by immunoblotting using AF and HSP90 antibodies. c–e Subcellular distribution of KLHL17/AF in mature neurons. Cultured hippocampal neurons were co-transfected with a Myc-tagged AF and GFP-actin construct at 12 DIV and immunostained at 18 DIV. c, d Confocal images. c Immunoreactivity of KLHL17/AF, as revealed by Myc antibody, shows punctate signal in the soma and dendrites of mature cultured neurons. d Locally enlarged dendrite (white box in c) to highlight the distribution of KLHL17/AF in dendrites and dendritic spines. Dendrite morphology was outlined by the signals of GFP-actin. Arrows indicate AF localized in the dendritic spine. Arrowheads indicate AF localized at the base of dendritic spines. e Super-resolution images of KLHL17/AF and F-actin immunoreactivities in a mature cultured neuron. AF is colocalized with or adjacent to F-actin at dendritic spines. F-actin signal was revealed by phalloidin-Alexa Fluor-405. Scale bars: c 20 μm; d 2 μm; e whole cell, 10 μm; enlarged dendrite segment, 2 μm; enlarged dendritic spine, 1 μm