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. 2018 Jan 17;115(5):998–1003. doi: 10.1073/pnas.1705954115

Fig. 1.

Fig. 1.

FBXO31 is regulated through proteasomal degradation promoted by the APC/C coactivators CDH1 and CDC20. (A) Immunoblot monitoring FBXO31, cyclin A, cyclin B, CDH1, CDC20, and phospho-histone H3 (p-H3) in HEK293T cells following HU synchronization and release. p-H3 was used as a marker for mitotic cells. α-Tubulin (TUBA) was monitored as loading control. (B) Immunoblot analysis monitoring the levels of FBXO31 in HEK293T cells expressing a NS shRNA or one of two unrelated APC2 shRNAs. (C and D) Immunoblot monitoring FBXO31 and cyclin B levels in HEK293T cells expressing vector or HA-CDH1 (C) or HA-CDC20 (D) in the presence or absence of MG132. β-Actin (ACTB) was monitored as a loading control. (E) Immunoblot monitoring FBXO31 levels in HEK293T cells expressing vector, HA-CDH1, or HA-CDC20 and either a NS or a APC2 shRNA. (F and G) Quantification of cycloheximide chase/immunoblot assay monitoring FBXO31 stability in HEK293T cells expressing vector, HA-CDH1 (F), or HA-CDC20 (G). (H and I) Immunoblot monitoring FBXO31 levels in HEK293T cells expressing a NS shRNA or one of two unrelated CDH1 (H) or CDC20 (I) shRNAs. (J and K) Quantification of cycloheximide chase/immunoblot assay monitoring FBXO31 stability in HEK293T cells expressing a NS, CDH1 (J), or CDC20 (K) shRNA. Data are represented as mean ± SD.