Fig. 5: FBXO47 maintains centromere stability by sustaining SCF complex in spermatocytes.
a, a1–3 WB analysis of CENP-C, SYCP1, and SKP1 expression in leptotene/zygotene cells and pachytene cells from both genotypes, as well as the quantitation of the results at pachytene (like) stage. b, b′ Location of SKP1 at synapsed regions of zygotene in Fbxo47+/+ spermatocytes. Arrows highlight the synapsed regions. c, c′ SKP1 location in pachytene spermatocytes of Fbxo47+/+ mice. Arrows indicate the synapsed regions. d, d′ Normal SKP1 signal observed in Fbxo47−⁄− zygotene cells. e, e′ Weakened SKP1 signal in pachytene-like cells of Fbxo47−⁄− mice. f, f′ SKP1 locates at centromeres on Fbxo47+/+ chromosomes. Arrows point to the centromeres, and insets show enlarged views. g, g′ Weak SKP1 signal in Fbxo47−⁄− pachytene-like cells. Scale bar, 5 µm. h Co-immunoprecipitation (Co-IP) study demonstrating that CENP-C1 can pull down overexpressed FBXO47 protein from HEK293T cell lysates. i SKP1 is also capable of pulling down overexpressed FBXO47 protein from HEK293T cell lysates. The arrow indicates Flag-SKP1. j Overexpression of FBXO47 leads to a decrease in the ubiquitination of SKP1 expressed in HEK293T cells. The arrow points to ubiquitinated Flag-SKP1. The Co-IP experiments were repeated three times with consistent outcomes. The data presented in this figure underscore the role of FBXO47 in maintaining centromere stability by ensuring the proper expression of SKP1 through the UPS pathway in spermatocytes.
