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. 2014 Aug 6;5:4619. doi: 10.1038/ncomms5619

Figure 8. Stiff matrix inhibits self-renewal of TRCs.

Figure 8

Stiff fibrin gels inhibit colony growth (a) and number (b). ‘90-Pa to 90-Pa’ and ‘90-Pa to 1,050-Pa’: control B16-F1 cells were cultured in 90-Pa fibrin gels for 5 days and these cells were then re-plated into 90-Pa and 1,050-Pa fibrin gels, respectively. ‘1,050-Pa to 90-Pa’ and ‘1,050-Pa to 1050-Pa’: control B16-F1 cells were cultured in 1050-Pa fibrin gels for 5 days and these cells were then re-plated into 90-Pa or 1,050-Pa fibrin gels. Spheroid size and number were measured from Day 1 through Day 5. In a and b, significant differences between ‘90-Pa to 90-Pa’ and ‘90-Pa to 1,050-Pa’ from Day 1 through Day 5 (all P<0.01); significant differences between ‘1,050-to 90-Pa’ and ‘1,050-Pa to 1050-Pa’ at day 4 and day 5 in a (both P<0.02) and from day 1 to day 4 in b (all P<0.05). Mean±s.e.m.; n=2 independent experiments. Each data point was averaged from >30 colonies. ‘*’ and ‘#’ indicates significant differences between ‘90-Pa to 90-Pa’ and ‘90-Pa to 1,050-Pa’ and between ‘1,050-to 90-Pa’ and ‘1,050-Pa to 1050-Pa’, respectively. (c) Switching to 3D soft matrix decreases H3K9 methylation. Mean±s.e.m.; n>50 cells from two independent experiments; *P<0.05. (d) Switching to 3D soft matrix increases Sox2 expression. Mean±s.e.m.; n=3 independent experiments; *P<0.05. In c and d, control cells were cultured in 1,050-Pa fibrin gels for 5 days and then re-plated into 90-Pa or 1,050-Pa gels. After 24 or 48 h in c and 5 days in d, their H3K9 methylation was measured by FRET and Sox2 expression was quantified by real time RT–PCR.