Additional file 3.
Figure S2: Cyclin B is detectable in nocodazole blocked cells. (A) Nocodazole blocked HeLa cells were visualized for Mcm10 and cyclin B. Mcm10 was visualized by anti-Mcm10 antibody in conjugation with anti-rabbit Alexa 488 antibody (top-right panel) while cyclin B was visualized with anti-cyclin B antibody in conjugation with anti-rabbit Alexa 488 antibody (bottom-right panel). DNA was stained with DAPI (left panel). HeLa cells that are blocked in mitosis have been marked by arrowheads. (B) HeLa cells were harvested after release from (Noc) and treated with MG132 as indicated and immunoblotted with anti-Mcm10 antibody (top panel). ASN refers to samples from asynchronously growing cells not treated with nocodazole while NS points to a non-specific band that displays equal protein loading. U2OS cells expressing HA-tagged full length Mcm10 (second panel) or HA-tagged linker domain of Mcm10 (third panel) were treated similarly. Since MG132 interfered with the nocodazole block in U2OS cells, the total population of cells after treatment with nocodazole and MG132 was loaded in second and third panel. The observed mobility of the linker domain of Mcm10 (fused to GFP and HA epitope tags) was approximately 55 kDa.
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Kaur et al. BMC Cell Biology 2010 11:84 doi:10.1186/1471-2121-11-84