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Fig. 5.

Analysis of -LacI-GFP and SPBs during mitosis in live wild type, and cells. Images from time lapse analysis of live cells showing the location of -LacI-GFP (green) and the SPB marker Cut12-CFP (red) in (A) wild-type (B) and (C) cells. Scale bar: 0.2 μm. The white and red arrows indicate the positions of the SPB Cut12-CFP signals (red arrows indicate inferred SPB positions from other images in the time course series) and the yellow arrows indicate the position of -LacI-GFP in cases where labeling was weak. The cytoplasmic red signal is nonspecific and due to autofluorescence detected with the CFP filter. Statistical analysis of anaphase -LacI-GFP movements in live wild-type, and cells is shown in Table 2 .

View this table:
Table 2.

Characteristics of anaphase movements in live wild-type, and cells

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K.E. is a Royal Swedish Academy of Sciences Research Fellow supported by a grant from the Knut and Alice Wallenberg Foundation. H.A. was the recipient of a VR-NT post doctoral grant. We thank R. Silverstein for critical reading of the manuscript. We thank M. Yanagida and Y. Hiraoka and I. Hagan for the generously providing us with strains. We thank K. Hultenby and S. Mengarelli (Huddinge Hospital, EM unit) for technical assistance with HPF and EM. This project was supported by Strategic (SSF-JIG) and Medical Research Council (VR-M 31X-12562) grants to K.E.

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Present address: Alfred Nobel's Alle 7, S-141 89, Huddinge, Sweden

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Allshire, R. C., Nimmo, E. R., Ekwall, K., Javerzat, J. P. and Cranston, G.
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Abstract / FREE Full Text Google Scholar
Bahler, J., Wu, J. Q., Longtine, M. S., Shah, N. G., McKenzie, A., 3rd, Steever, A. B., Wach, A., Philippsen, P. and Pringle, J. R.

Importantly, healthy subjects and patients with type 1 diabetes who received insulin glargine injections showed free serum insulin glargine levels between 70 to 90 pmol/L with peak levels of about <200 pmol/L after injection [52] Rick Owens Black amp; OffWhite Geothrasher Low Sneakers J6zPO0HXCs
. The therapeutic insulin glargine concentrations reported so far are 80- to 200-fold below the total plasma concentrations of IGF-1. Notably, up to 99% of the circulating IGF-1 is bound to the IGF-binding proteins 1–6 in human plasma [13] , Converse Womens Chuck Taylor All Star Shoreline Peached Twill Sneaker L9omZgJdk
. Therefore, the concentration of free IGF-1 may be within the range of the plasma concentration of insulin glargine. However, despite the putative comparable plasma concentrations the ∼100-fold reduced affinity of insulin glargine towards IGF1R compared to that of IGF-1 makes a competition even of non-metabolized insulin glargine with endogenous IGF-1 at the receptor unlikely. Furthermore, the concentration of free IGF-1 may be elevated during certain diseases and metabolic states, which are known to considerably reduce the expression of IGF-binding proteins [57] , Nike Mens Kyrie Flytrap Basketball Sneakers from Finish Line ucZmqSKQ
making interaction of insulin glargine with the IGF1R in course of displacement of IGF-1 even less likely.

Insulin glargine has been evaluated extensively in a 2-year life-time carcinogenicity study in rats without evidence of tumor-promoting activity greater than human insulin [59] . In these studies plasma insulin concentrations were 15 to 25 nmol/L, covering parent compound and active metabolites. In consequence, actual glargine concentrations may be overestimated, the extent of which depends on the degree of proteolytic degradation at both the injection site and the serum. Quantitative assessment of the insulin glargine metabolism in humans is evolving, and will bring further clarification.

Conclusion

The present data unequivocally demonstrate similar metabolic signaling and activity in rat adipocytes of insulin glargine and its major metabolites with comparable engagement of IR-A and IR-B. In contrast, the insulin glargine metabolites were found to be less potent in mitogenic signaling and less active in Saos-2 proliferation assay compared to insulin glargine. The formation of insulin glargine metabolites may help to explain the established efficacy and safety of insulin glargine as observed in clinical practice and particularly in long-term clinical trials.

Acknowledgments

We thank Niels Griesang for synthesis of iodized human insulin and Kathi Klein, Gerhard Hoffmann, Stefan Welte, Susanne Wied for excellent technical assistance.

Author Contributions

Conceived and designed the experiments: MRS GM GT NT. Performed the experiments: MRS GM GT NT. Analyzed the data: MRS GM GT NT. Contributed reagents/materials/analysis tools: GS PH. Wrote the paper: MRS GM GT NT. Revised the manuscript for important intellectual content: RK.

References

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