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. Author manuscript; available in PMC: 2015 Dec 22.
Published in final edited form as: Dev Cell. 2014 Dec 11;31(6):722–733. doi: 10.1016/j.devcel.2014.11.012

Figure 5. PDGF-A Is Expressed and Secreted by Senescent Cells.

Figure 5

(A) mRNA levels of the indicated proteins were quantified by qRT-PCR in cells sorted from wounds as described in Figure 4D. Actin was used to control for RNA quantity (n = 3).

(B) Skin biopsies excised from PBS- or GCV-treated wounds in p16-3MR mice were collected at the indicated intervals after injury, and PDGFA mRNA levels were quantified by qRT-PCR. Actin was used to control for RNA quantity (n = 5).

(C) Skin biopsies of p16-3MR mice were collected 6 days after injury, fixed, stained with DAPI (blue; indicating nuclei), and immunostained for mRFP (red) and PDGF-A (green).

(D) Cells were derived from wounds 6 days after injury and sorted for RFP. RFP+ cells were plated, fixed 24 hr later, and then stained with DAPI (blue) and immunostained for PDGF-A (red) and vimentin (green, left panels) or factor VIII (green, right panels).

(E and F) Murine skin fibroblasts (SF) or endothelial cells (EC) were mock irradiated (NS) or made senescent by irradiation (SEN; 10 Gy X-ray). At 4 days after irradiation, RNA and conditioned media were collected and analyzed for PDGF-A mRNA and secreted protein by qRT-PCR and ELISA, respectively (n = 4). Data shown are the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.0001.