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. 2016 Dec 8;7:13674. doi: 10.1038/ncomms13674

Figure 4. TXNIP-derived peptide (TAT-TN13) inhibits p38 activity and rejuvenates aged HSCs in vitro.

Figure 4

(a) Phosphorylation of p38 and ATF2 in old BM cells. Freshly isolated old BM cells were treated with TAT, TAT-TN13 or SB203580 for 1 h in RPMI 1640 containing 10% FBS (repeated two times). (b) Confocal images of phopho-p38 in old HSCs. Sorted old LT-HSCs were treated with 10 μM TAT or TAT-TN13 for 1 h in HSC media (repeated three times). (c,d) Immunoprecipitation assay in old BM cells (c) and in situ PLA images in old HSCs (d). Old BM cells and old LT-HSCs were treated with 10 μM TAT or TAT-TN13 for 1 h (repeated two or three times). (e,f) Levels of phospho-p38 (e) and ROS levels (f) in old HSCs. Sorted LT-HSCs were treated with 10 μM TAT, TAT-TN13 or SB203580 for 16 h in HSC media (n=3 from two experiments). (g) Polar distribution of Cdc42 in LT-HSCs. Sorted LT-HSCs were treated with 10 μM TAT, TAT-TN13 or SB203580 for 16 h in HSC media (repeated two times). (h–k) Quantitative real-time PCR of ageing-associated genes in old HSCs. Sorted LT-HSCs were treated with 10 μM TAT, TAT-TN13 or SB203580 for 16 h in HSC media (n=3 from two experiments). (l) The homing ability of old HSCs. Sorted LT-HSCs were treated with 10 μM TAT, TAT-TN13 or SB203580 for 16 h in HSC media (n=6–10). Data are mean±s.d. Statistical significance was determined using a two-tailed Student's t-tests. *P<0.05, **P<0.01, ***P<0.001.