Figure 1.Depletion of TSC2 converts quiescence into senescence in HT-p21-9 cells.(A) HT-p21-9 cells
were transduced with control lentivirus (pLKO) or lentivirus expressing
shTSC2 (sequence # 10) and selected with puromycin for 5 days and then
immunoblot was performed. (B)
Schema: Testing the reversibility of nutlin-3a effects. (C) HT-p21-9 cells were transduced
with control pLKO or shTSC2 and 5000 cells were plated in 24-well plates
and, the next day, were treated with 10 uM nutlin-3a for 3 days. Then
nutlin-3a was washed out and the cells were cultivated in fresh medium for
3 days and then stained for beta-Gal and microphotographed. Bars 50 um. (D) HT-p21-9 cells were transduced
with control pLKO or shTSC2 (and selected for 4 days with puromycin). Then
1000 cells were plated per 60-mm dishes and, the next day, were treated with
nutlin-3a for 3 days. Then nutlin-3a was washed out and cells were
cultivated in fresh medium for 8 days. Colonies were stained with crystal
violet.
