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. 2020 Oct 8;39(49):7166–7180. doi: 10.1038/s41388-020-01488-5

Fig. 5. SHP2 facilitates signaling in response to several growth factors.

Fig. 5

a Immunoblot analyses showing the phosphorylation of SHP2 and ERK1/2 (ERK) induced by the indicated growth factors in D2.A1 cells. D2.A1 cell growth was induced by addition of exogenous FGF2, PDGF, and hGF in 2D (b) or 3D (c) culture for 6 days. D2.A1 cells expressing doxycycline inducible shRNAs targeting PTPN11 were treated with doxycycline in 2D culture (d) or 3D culture (e) in the presence of FGF2, hGF, or PDGF as indicated. For b–e cell viability was quantified as a relative bioluminescence ratio normalized to day 0. Data are the mean ± s.e.m. for at least three independent assays resulting in *p < 0.05, **p < 0.01, ***p < 0.001, or no significance (NS) as determined by a two-tail Student’s t test. f–h D2.A1 cells were pre-treated with SHP099 or FIIN4 for 24 h in serum-free media and cells were subsequently induced for 5 min with FGF2, PDGF, or hGF as indicated. Immunoblot analyses were used to detect phosphorylation of FGFR, FRS2, ERK1/2, AKT, and SHP2. i D2.A1 cells were pre-treated with PP2 or PF271 for 24 h in serum-free media, and cells were then induced for 5 min with FGF2, PDGF, or hGF. Immunoblot analyses were used to detect phosphorylation of SHP2. All immunoblots are representative of at least three independent experiments.