Figure 4.
Mut-KIF4 strongly binds to PARP1, whose activity influences cell survival rate. (A and B) Co-immunoprecipitation (Co-IP) immunoblotting results of the primary culture neuron lysates (A), and quantification of amounts of 110 kD PARP1 bands (B). The statistical values were normalized to relevant bands in the input lanes. The input samples and immunoprecipitated complexes including KIF4 were analyzed by immunoblotting using an antibody specific to PARP1; IP, immunoprecipitation; IB, immunoblotting; IgG, immunoglobulin G; Cyt., Cytosol; Nuc., Nuclear. Data are presented as mean ± SEM (n = 3 independent experiments). ***P < 0.001 (Student’s t tests). (C and D) Immunoblotting results of BioID pulldown (C), and quantification of amounts of PARP1 protein bands (D). The statistical values were normalized to overexpressed KIF4 bands. The biotinylated PARP1 was clearly detected in a lysate of N2A cells expressing Mut-KIF4. Data are presented as mean ± SEM (n = 3 independent experiments). ***P < 0.001 (Student’s t tests). (E and F) The survival assay using CMFDA with or without indicated reagents. Representative images of hippocampal culture neurons (E), and quantification of survival rates for each condition (F). Scale bar, 50 µm. Data are presented as mean ± SEM (n = 3 independent experiments). ns, P > 0.05, ****P < 0.0001 (two-way ANOVA). (G and H) Hippocampal neurons immunostained with anti-PAR and anti-MAP2 antibodies (G), and quantification of the normalized fluorescent intensity for each condition (H). Scale bar, 10 µm. Data are presented as mean ± SEM (n = 60, 3 independent experiments). ns, P > 0.05, ****P < 0.0001 (Student’s t tests). Source data are available for this figure: SourceData F4.

Mut-KIF4 strongly binds to PARP1, whose activity influences cell survival rate. (A and B) Co-immunoprecipitation (Co-IP) immunoblotting results of the primary culture neuron lysates (A), and quantification of amounts of 110 kD PARP1 bands (B). The statistical values were normalized to relevant bands in the input lanes. The input samples and immunoprecipitated complexes including KIF4 were analyzed by immunoblotting using an antibody specific to PARP1; IP, immunoprecipitation; IB, immunoblotting; IgG, immunoglobulin G; Cyt., Cytosol; Nuc., Nuclear. Data are presented as mean ± SEM (n = 3 independent experiments). ***P < 0.001 (Student’s t tests). (C and D) Immunoblotting results of BioID pulldown (C), and quantification of amounts of PARP1 protein bands (D). The statistical values were normalized to overexpressed KIF4 bands. The biotinylated PARP1 was clearly detected in a lysate of N2A cells expressing Mut-KIF4. Data are presented as mean ± SEM (n = 3 independent experiments). ***P < 0.001 (Student’s t tests). (E and F) The survival assay using CMFDA with or without indicated reagents. Representative images of hippocampal culture neurons (E), and quantification of survival rates for each condition (F). Scale bar, 50 µm. Data are presented as mean ± SEM (n = 3 independent experiments). ns, P > 0.05, ****P < 0.0001 (two-way ANOVA). (G and H) Hippocampal neurons immunostained with anti-PAR and anti-MAP2 antibodies (G), and quantification of the normalized fluorescent intensity for each condition (H). Scale bar, 10 µm. Data are presented as mean ± SEM (n = 60, 3 independent experiments). ns, P > 0.05, ****P < 0.0001 (Student’s t tests). Source data are available for this figure: SourceData F4.

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