Fig. 7.

Cellular senescence can be transmitted in vitro. Inhibition of TGFβ decreases paracrine senescence. a Experimental scheme; biliary cells were treated with RAS-conditional media (CM-RAS) and CONTROL-conditional media (CM-Ctrol) for a week. Media is eliminated and cells washed with PBS five times. YFP-positive biliary cells (YFP-biliary cells) are then added and after 1 week different markers of senescence in the YFP-biliary cells population were assessed. b Representative images of the assessed markers of senescence and proliferation (BrdU) in the co-culture of senescent biliary cells (YFP-negative) and YFP-positive cells. Data is presented as biliary cells treated with CM-STOP and CM-RAS at final time point (14 days). Far right, percentage of each marker in the YFP-biliary cells population. * denotes p < 0.05 (Mean ± SEM). Mann−Whitney test. N = 4 biological replicates. Scale bars = 50 µm. c Experimental scheme for the use of TGFβ inhibitor (LY-2157299); YFP-biliary cells and different concentrations of LY-2157299 are added to the senescent biliary cells. After 1 week different markers of senescence in the YFP-biliary cells population were assessed. d Decrease of senescence factors and increase in proliferation in YFP-biliary cells with increasing concentrations of LY-2157299. * denotes p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (Mean ± SEM). ANOVA, Sidak’s multiple comparisons test. N = 4 biological replicates