Intracellular localization of KCTD20 or expression level of KCTD20 in mouse spinal cord anterior horn. A, His-Xpress-KCTD20 was expressed in COS7 cells by transfection of pEF4-KCTD20 endocing HisXpress-tagged KCTD20. The backbone pEF4 vector was similarly transfected as a negative control. Transfected cells were fixed with 4% paraformaldehyde and immunostained with anti-Xpress antibody as a primary antibody and FITC-conjugated anti-mouse IgG antibody as a secondary antibody. The scale bar indicates 20 μm. B, COS7 cells were transfected with pEF4-KCTD20 and pCAGGS-BTBD10 and fixed at 48 hr after transfection. The cells were immunostained with Xpress or BTBD10 antibody as a primary antibody and FITC-conjugated anti-mouse IgG antibody or TexasRed-conjugated anti-rabbit IgG antibody as a secondary antibody, respectively. C, Frozen sections of spinal cords of G93A-SOD1-Tg mouse or wild-type littermate were immunostained with KCTD20 antibody. Rightmost pictures in each series (indicated as day 120 preabsorption) were immunostained using KCTD20 antibody, preabsorbed with the antigen peptide. Areas, surrounded by dashed lines, represent spinal ventral horns. The scale bar indicates 50 μm.