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

Figure 2. FGFR-activated MAPK signaling depends on SHP2.

Figure 2

Immunoblot of indicated proteins in JHH-7 cells transfected with siRNAs targeting SHP2 or FRS2 or scrambled control siRNA for 3 days (A) or Hep3B and JHH-7 cells stably expressing doxycycline-inducible shRNA targeting SHP2 (shSHP2) treated with PBS (-) or 100 ng/ml doxycycline (+) for 4 days (B). (C) Colony formation assay with cells described in (B) treated with DMSO, 100 ng/ml doxycycline (shSHP2), 0.1 μM BGJ398, or 10 μM SHP099 for 10–14 days, followed by crystal violet staining. (D) Immunoblot of SHP2 and tubulin with cells described in (B) that were further engineered to constitutively express shSHP2-resistant SHP2 (with a biotin ligase tag, upper band) and parental JHH-7 or Hep3B cells as a control (first lane). (E) Colony formation assay with cells described in (D) treated with DMSO, 100 ng/ml doxycycline (+shSHP2), or doxycycline plus 0.1 μM BGJ398 for 14–21 days, followed by crystal violet staining.