Inactivation of p16INK4a (inhibitor of cyclin-dependent kinase 4A) Immortalizes Primary Human Keratinocytes By Maintaining Cells in the Stem Cell Compartment FASEB J. Maurelli et al.
20: 1516
Supplemental Data
Files in this Data Supplement:
Supplemental Figure 1
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(SupplementalFig1.JPG; 17 KB)
TRF assay performed on cells transduced with empty vector (V lanes) and antisense p16INK4a
-transduced cells (AS-Exo1a lanes) at different passages (indicated by the numbers of cell generations) after infection.
Supplemental Figure 2
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(SupplementalFig2.JPG; 29 KB)
hTERT expression was evaluated by quantitative RT-PCR analysis on RNA extracted from empty vector (column 1) and sense p16INK4a (column 2) transduced keratinocytes, antisense p16INK4a-transduced keratinocytes infected with empty vector (column 3) and antisense Bmi-1 (column 4). RNA from Jurkat cells (column 5) was used as positive control of hTERT expression. The RNA levels were normalized using the GA3PDH gene as housekeeping gene. Results were expressed as relative levels of hTERT mRNA referred to a calibrator sample, the empty vector control (column 1), which was chosen to represent 1x expression of this gene. The error bars represent standard deviation of triplicate experiments. The axis break is between 0.025 and 1.
Supplemental Figure 3
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(SupplementalFig3.JPG; 344 KB)
Immunostaining of Bmi-1 was performed on normal human keratinocyte cells (A), antisense p16INK4a-transduced cells (B) and antisense p16INK4a-antisense-Bmi-1-transduced cells (C). Black and red arrows label nuclear and cytoplasmic staining, respectively.
