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Figure 5


Fig. 5. Keratinocyte differentiation is accompanied by a transient upregulation of several sumoylation system proteins. Parallel HaCaT cell cultures were maintained either in low [Ca2+] medium (Uninduced) or switched from low [Ca2+] to high [Ca2+] medium (Induced). At the indicated times post Ca2+ induction, protein extracts were prepared from the paired cultures. The extracts were assayed for expression of K1 (A), Ubc9 (B), SAE1 (B), and SENP1 (B) by immunoblotting with specific antibodies. The induced and uninduced cell extracts were also evaluated for {alpha}-tubulin expression, and the immunoblots for {alpha}-tubulin are duplicated in each panel of B for comparative purposes. (C) The immunoblots in B were quantified by densitometry, and the values for Ubc9, SAE1, and SENP1 were normalized against {alpha}-tubulin. The 0 hour time values for both the induced and uninduced samples were set to 1, and the values for the later time points are relative to the 0 hour time. The quantitative results in C are the average of at least three independent experiments.





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