Interaction of CCNYL1 with CDK16.

<p>(A) Co-immunolabeling of CCNYL1 (red) and CDK16 (green) in testicular sections of WT mice. Nuclei were labeled with DAPI (blue). Scale bar: 50 μm. (B) CDK16 protein expression was measured in membrane extracts and cytoplasmic extracts isolated from testes or germ cells of WT and <i>Ccnyl1-/-</i> mice. N-cadherin served as the membrane protein control, while β-actin served as the cytoplasmic protein control. (C) CDK16 protein was immunoprecipitated from testicular lysate with CDK16 antibody. Both CDK16 and the Co-immunoprecipitated CCNYL1 were analyzed by western blotting. (D) HEK293T cells were co-transfected with N-terminal tagged CCNYL1<i>-</i>HA and C-terminal tagged CDK16-Flag plasmids. CCNYL1-HA and CCNYL1-Flag proteins were co-immunoprecipitated using either anti-HA or anti-Flag conjugated agarose before analyzed by western blot. (E) HEK293T cells were co-transfected with CDK16-Flag and CCNYL1-HA, or CDK16-Flag and CCNYL-HA plasmids, proteins were co-immunoprecipitated by using anti-Flag conjugated agarose before analyzed by western blotting. (F) Truncated CCNYL1-HA variants were coexpressed with CDK16-Flag in HEK293T cells. The interactions were analyzed by CoIP experiments followed by western blotting. (G) CDK16-Flag variants were co-expressed with CCNYL1-HA in HEK293T cells. The interactions were analyzed by CoIP experiments followed by western blotting. PCTAIRE is the conserved sequence of CDK16 protein. PCTAIRE mutant (PCTAIRE was mutated to ACAAIAA).