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. 2020 Jan 21;11(3):265–281. doi: 10.18632/oncotarget.27435

Figure 5. Rapid FGFR feedback activation is caused by downregulation of Sprouty proteins.

Figure 5

(A) mRNA levels determined by qRT-PCR of SPRY1, SPRY2, and SPRY4 in Detroit 562, KYSE-520, Hep3B, and JHH-7 cells treated with DMSO, 3 μM SHP394, 0.5 μM RTK inhibitor (erlotinib for Detroit 562 and KYSE-520, BGJ398 for Hep3B and JHH-7), or 10 nM trametinib for 2 h. Statistical analysis (paired t-tests) was described in Supplementary Table 3. (B) Hep3B cells stably expressing doxycycline-inducible SPRY4 were treated with PBS (−Dox) or 100 ng/ml doxycycline (+SPRY4) for 48 h, followed by treatment with DMSO, SHP394 at indicated concentrations, or 0.5 μM BGJ398 for either 15 min or 2 h. Cell lysates were collected and p-FRS2 (Y436), p-SHP2 (Y542), p-ERK1/2, and SPRY4 levels were determined by immunoblotting. (C) Anti-proliferative effects of SHP394 at the indicated concentrations in a 6-day cell proliferation assay with Hep3B and JHH-7 cells stably expressing doxycycline-inducible SPRY4 in the absence (−Dox) or presence (+Dox/SPRY4) of 100 ng/ml doxycycline (mean percentages of cell viability are shown; error bars, SD; n = 3).