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Tissue-Specific Stem Cells
Version of Record online: 22 OCT 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 11, pages 2512–2522, November 2012
How to Cite
Cottage, C. T., Neidig, L., Sundararaman, B., Din, S., Joyo, A. Y., Bailey, B., Gude, N., Hariharan, N. and Sussman, M. A. (2012), Increased Mitotic Rate Coincident with Transient Telomere Lengthening Resulting from Pim-1 Overexpression in Cardiac Progenitor Cells. STEM CELLS, 30: 2512–2522. doi: 10.1002/stem.1211
Author contributions: C.T.C.: conception and design, collection and/or assembly of data, data analysis and interpretation, and manuscript writing, L.N., B.S., S.D., A.J., B.B., N.G., and N.H.: collection and/or assembly of data, M.S.: conception and design, financial support, manuscript writing, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS August 22, 2012.
- Issue online: 22 OCT 2012
- Version of Record online: 22 OCT 2012
- Accepted manuscript online: 22 AUG 2012 10:40AM EST
- Manuscript Accepted: 23 JUL 2012
- Manuscript Received: 7 JUN 2012
- National Heart, Lung, and Blood Institute. Grant Numbers: R21-HL102714, R01-HL067245, R37-HL091102, P01-HL085577, RC1-HL100891, R21-HL102613, R21 HL104544, R01 HL105759
- Rees-Stealy Research Foundation
- San Diego ARCS Foundation
- American Heart Association Pre-doctoral Fellowship. Grant Number: 10PRE3060046
- Inamori Foundation Fellowship
Additional Supporting Information may be found in the online version of this article.
|sc-12-0544_sm_SupplTable1.pdf||47K||Supplementary Table 1|
|sc-12-0544_sm_SupplFigure1.tif||495K||Supplemental Figure 1. Increased population cycling in early passage CPCeP and CPCs isolated from 13 month old hearts and infected with Pim-1, related to Figure 1. A. Cyquant assay performed on CPCe and CPCeP passage 5 (P5) and measured at 4, 6, 24, and 30 hours after plating. B. Cyquant assay performed on isolated older CPCs infected with lentivirus overexpressing GFP (OCPCe) or Pim-1 (OCPCeP). #p=0.09 vs. OCPCe day 3, *p<0.05 vs. OCPCe day 5.|
|sc-12-0544_sm_SupplFigure2.tif||861K||Supplemental Figure 2. Western blots from quantitations in text, related to Figures 2 and 4. A-C. Immunoblots of CPCe and CPCeP at varying passages and probed using antibodies against the indicated proteins. + indicates whole lysates from Hela cells, e is CPCe and eP is CPCeP. ++ Indicates whole cell lysates from CPCs infected with lentivirus overexpressing TERT|
|sc-12-0544_sm_SupplFigure3.tif||67K||Supplement Figure 3. Telomere lengthening in CPCeP is dependent on TERT activity, related to Figure 3. A-D. Early passage CPCs treated with either DMSO or telomerase inhibitor (MST-312, 1μM).|
|sc-12-0544_sm_SupplFigure4.tif||154K||Supplemental Figure 4. Pim-1 interacts with TERT in CPCePs, related to Figure 5. A. Pim-1 and TERT interactions demonstrated by PLA for Pim-1 and TERT (red). B. Coimmunoprecipitation of Pim-1 and TERT. C PLA performed on CPCeP without primary antibodies; this control was performed with every assay to confirm specific staining. D and E. PLAs of Pim-1 and TERT (red), Pim-1 and pS/pT and Tert (red) in CPCeP treated with Pim-1 inhibitor, Quercetagetin (Q, 10μM for 24 hours). F. Pim-1 expression knocked down with a Pim-1 specific siRNA. Lane 1 is CPCeP, lane 2 is CPCe+ siPim- 1, and lane 3 is CPCeP +siPim-1. G and H. PLAs of Pim-1 and TERT (red) and pS/pT and Tert CPCeP treated with siPim-1.|
|sc-12-0544_sm_SupplFigure5.tif||237K||Supplemental Figure 5. TERT expression and DNA incorporation after Dox treatment, related to Figure 7. A. Immunoblots of CPCe and CPCeP treated with Dox and probed for indicated antibodies + indicates whole cell Hela lysates. B. DNA synthesis measured by BrdU incorporation in DOX treated and untreated CPCs. $ p<0.05 vs untreated CPCe.|
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