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. 2016 Apr 20;7(19):26935–26948. doi: 10.18632/oncotarget.8854

Figure 2. Premature senescence associated with Gadd45b deficiency can be rescued, in part, by culture at low oxygen.

Figure 2

A. Gadd45b+/+ (WT - black), and Gadd45b−/− (KO - grey) MEFs were cultured at either 21% oxygen (square) or 3% oxygen (circle) for 14 passages. Cells were split every 3 days, and the total numbers of cells were counted and mean population doublings (MPD) were determined. B. Cumulative cell number is plotted against passage number. C. Gadd45b+/+ (black) and Gadd45b−/− (grey) MEFs, cultured at either 21% oxygen (solid) or 3% oxygen (dotted) were stained for SA-β-gal at each passage. SA-β-gal positive cells were counted in at least 10 fields from triplicate plates. A quantification of SA-β-gal positive Gadd45b+/+ and Gadd45b−/− MEFs is shown for each passage. D. Gadd45b+/+ MEFs were cultured at either 21% oxygen or 3% oxygen. Total RNA for each passage was analyzed by real time PCR for Gadd45b expression using taqman probe as described in Materials and methods. 18S rRNA probe was used as an internal control.