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. Author manuscript; available in PMC: 2014 Mar 1.
Published in final edited form as: J Neuropathol Exp Neurol. 2013 Mar;72(3):219–233. doi: 10.1097/NEN.0b013e3182859939

Figure 2.

Figure 2

Epidermal growth factor receptors (EGFRs) are required for the migration of unstimulated malignant peripheral nerve sheath tumor (MPNST) cells. (A) ST88-14 MPNST cells stably transduced with a pSLIK lentivirus coordinately expressing an EGFR shRNA and green fluorescent protein (GFP) were treated with a range of doxycycline concentrations for 72 hours. These lysates were then probed for EGFR (upper panel) and GFP (30 kDa; middle panel) to assess EGFR knockdown and cassette activation, respectively. The lower panel is a glyceraldehyde-3-phosphate dehydrogenase (GAPDH) loading control. (B) Migration of ST88-14 MPNST cells 72 hours after inducing the expression of 2 distinct EGFR shRNAs (shEGFR1, shEGFR2) or a nonsense control shRNA. Migration in the EGFR shRNA-expressing cells was normalized to migration in cells expressing the nonsense control shRNA. SEM are indicated for each condition; *p < 0.05 vs. the nonsense control. (C) T265-2c MPNST cell lysates were prepared 72 hours after transfection with plasmids expressing 2 distinct EGFR shRNAs (shEGFR3, shEGFR4) or a nonsense control shRNA and probed for EGFR expression to demonstrate knockdown (upper panel). The lower panel is a GAPDH loading control. (D) Migration assays with T265-2c MPNST cells transiently transfected with the same shRNA-expressing plasmids expressing shRNAs presented in (C). Numbers of shEGFR transfected cells migrating to filter undersurfaces are normalized to numbers of nonsense shRNA transfected cells migrating to filter undersurfaces under the same conditions. SEM are indicated, with asterisk (*) indicating conditions for which p < 0.05 for comparisons with the nonsense control.