Research Paper Volume 14, Issue 10 pp 4281—4304

Figure 2. Cell senescence increases with age in AhR-deficient liver. (A) SA-β-Gal activity was analyzed in AhR+/+ and AhR−/− liver sections by staining with the chromogenic substrate X-Gal. (B) SA-β-Gal activity was analyzed by FACS in isolated liver cells (gentleMACS) using the fluorescent substrate C12FDG. (C–F) mRNA levels of senescence driver genes p16^Ink4a (C) and p21^Cip1 (D) and SASP-related genes IL1 (E) and TNFα (F) were analyzed by RT-qPCR in AhR+/+ and AhR−/− livers at the indicated ages. Oligonucleotides used are indicated in Supplementary Table 1. Gapdh was used to normalize target gene expression (△Ct) and 2^−△△Ct to calculate changes in mRNA levels with respect to wild type or untreated conditions. Data are shown as mean + SD (^*P < 0.05; ^**P < 0.01; ^***P < 0.001).