Cellular senescence is induced by TSLP stimulation <i>in</i><i>vitro</i>.

<p>(<b>A</b>) TSLP-induced p16 and p21 upregulation occurs in a TSLP dose-dependent manner in human bronchial epithelial BEAS-2B cells. BEAS-2B cells were stimulated with different doses of TSLP as indicated for 6h. Protein expressions of p16, p21 and phospho-Stat3 (Try705) were detected by western blotting. (<b>B</b>) BEAS-2B cells were stimulated by 1.5ng/ml TSLP and protein expressions of p16 and p21 were detected by western blotting. (<b>C</b>) BEAS-2B cells were stimulated with 1.5ng/ml TSLP then stained for BrdU. (<i>*p< 0.05</i>). BEAS-2B cells were stimulated with 1.5ng/ml TSLP then stained for SA-β-gal activity at 6 and 24 hours post stimulation. (<b>D</b>) upper panel: SA-β-gal staining; lower panel: quantification of SA-β-gal positive cells. (*<i>p</i> < 0.05); (<b>E</b>) Ki67 staining. (<b>F</b>) Levels of TSLP in culture media were examined by ELISA. </p

Experimental therapies with WP1066 in OVA-challenged chronic asthmatic mice.

<p>(<b>A</b>) OVA-challenge was performed in BALB/c mice as described [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077795#B28" target="_blank">28</a>]. WP1066 was administered by intraperitoneal injection at doses of 40mg/kg 1h before the OVA-challenge. Airway resistance was measured using increasing concentration of methacholine and assessed using the flexiVent system. Results are expressed as the mean of experiments done in triplicate ± the standard error of the mean (SEM) (<i>* p<0.05</i>, <i>**p>0.05 </i><i>vs </i><i>control</i>). (<b>B</b>) TSLP, p16, p21, Ki-67, α-SMA and collagen I protein expressions were analyzed, 200×. (<b>C</b>) Bimodal H score distribution of TSLP, p16, p21, Ki67, α-SMA and collagen I immunoperoxidase reactions are presented.</p

Senescent inhibition overcomes TSLP-induced airway remodeling in vitro.

<p>BEAS-2B cells with stable shp16, shp21 or both were incubated with TSLP (1.5ng/ml) for 6 h. (<b>A</b>) Cells were collected and total proteins were extracted and analyzed by western blotting. (<b>B</b>) Cells were fixed and stained with SA-β-gal (upper panel) and then positive SA-β-gal cells were quantified (<i>*p>0.05</i>, **<i>p</i> < 0.05) (low panel). (<b>C</b>) Cells were stained with BrdU (<i>*p>0.05</i>, **<i>p</i> < 0.05). (<b>D</b>) Senescent inhibition overcomes TSLP-induced cell growth inhibition in vitro. The relative cell number was detected to evaluate cell growth at different time points using MTT assays.</p