Following Scf treatment, robust pERK and pS6 responses were observed in LSK and LK, but not LDN cells (Figure 2A; quantified in Figure S3).
(A ) In the samples submitted for RNA-seq, both PDGF and FGF induce robust pERK responses at 15 minutes.
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(A ) Signaling time course shows a more robust phospho-ERK (pERK) response to FGF (blue) than PDGF (red) and a similar pAkt response to both growth factors.
In the samples submitted for sequencing, both PDGF and FGF induced a robust phospho-ERK (pERK) response at 15 min, and MEPMs generated from Pdgfra-GFP (Hamilton et al., 2003) and Fgfr1-CFP (to be described elsewhere) knockin reporter embryos display expression of each receptor at the protein level in all cells, further validating MEPMs as a suitable system to study RTK responses.
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For both the HSF1 and PERK inhibitors the most robust effects were seen at concentrations in excess of the in vitro IC50 (∼1.2 µM for KRIBB11, ∼1 nM for GSK2606414).
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In addition, a FGF-PKC-AP-1 signaling axis in calvarial osteoblasts has been reported (Miraoui et al., 2010), consistent with the MEPM data linking FGF to a robust AP-1 response and FGF-mediated pERK activation to PKC signaling.
