Determining the mechanosensitive targets of ERK and Wnt pathways in embryonic stem cells

Embryonic stem cells can be maintained in a state of naive pluripotency by the use of 2 inhibitors, PD03 - a MEK inhibitor upstream of ERK, and CHIRON, a GSK3 b inhibitor, which acts as a Wnt agonist. The removal of these inhibitors allow embryonic stem cells to exit pluripotency. To determine if targets of transcriptional targets of these pathways are mechanosensitive such that their activation or suppression would be dependent on mechanical properties of substrate. Overall design: Embryonic stem cells were cultured in pluripotency conditions 2i+LIF on soft or stiff StemBond hydrogels for 12 hrs. Then PD03 or CHIRON were removed for 2hrs and 12hrs before collecting RNA lysates for sequencing, and control samples remained in 2i+LIF served as 0hr timepoint . 2 replicates per condition were sequenced.