Fig. 6

Disruption of the SUN1 expression and location in Cdkn2aip^−/− mice. A Schematic representation of Bio-layer Interferometry (BLI). B Association and dissociation curves of CDKN2AIP protein combining with the ssDNA in a concentration range between 100 mM(blue), 150 mM(green), 200 mM(red). C A representative mRNA assembly output showing RIP-Seq reads for Sun1 identified from the RIP products using the CDKN2AIP antibody and Control. The red peak is the anti-CDKN2AIP bond and the bule peak is the control group. D qPCR analyses of level of CDKN2AIP-bound Sun1 mRNA in RIP products and Control. Data are presented as mean ± S.D. Student’s t test; **P < 0.01. E Validation of interactions between CDKN2AIP and SUN1 in protein level by co-immunoprecipitation assays, in which antibodies specific for CDKN2AIP was used for immunoprecipitation (IP) followed by western blot using anti-SUN1 antibody. IgG was used as a control. F Localization of SUN1 in the Cdkn2aip^+/+ and Cdkn2aip^−/− testis detected by immunohistochemistry. SUN1 is stained brown. Scale bar, 50 μm; Quantification of SUN1-positive signal in five horizons per group. Data are presented as mean ± S.D. Student’s t test; *P < 0.05. G Western blot analyses the expression of SUN1 in Cdkn2aip^+/+ and Cdkn2aip^−/− testis. GAPDH is serves as a loading control. H Immunostaining of SUN1(green) and SYCP3(red) on chromosome spreads of spermatocytes from P35 Cdkn2aip^+/+ and Cdkn2aip^−/− testes, n = 3 mice for each group. Scale bars, 10 μm. I Statistical analysis of the number of SUN1 positive signals in 10 pachytene spermatocytes from P35 Cdkn2aip^+/+ and Cdkn2aip^−/− testes. n = 3 mice for each group, and 10 pachytene spermatocytes were counted for each mouse. Data are presented as mean ± SD, **P < 0.01 by two-tailed Student’s-test