Figure 1.
Inhibiting the expression of Rab35 increases tumor growth in vivo. (A) BT025 cells were transduced with a nontargeting control (ctrl) shRNAmiR or two different shRNAmiRs targeting Rab35 (nt63 and nt419). Cell lysates were processed for immunoblot using antibodies recognizing the indicated proteins. The migration of molecular mass markers (in kD) is indicated. (B) BT025 cells (5 × 105 cells) transduced as in A were stereotactically injected into the right striatum of NOD-SCID mice, which were euthanized 1 wk after implantation, and brains were dissected. Tumor growth was monitored based on GFP fluorescence from the viral cassette. Three representative mouse brains (dorsal and ventral views) are shown for each condition. Scale bar, 2 mm. (C and D) Hematoxylin and eosin staining (C) and GFP staining (D) of coronal sections of brains of mice implanted with BT025 cells transduced with control (ctrl) or Rab35 nt63 shRNAmiRs. Scale bars, 2 mm. Arrows point to the tumors. Magnified views of the tumors in D are shown as insets with the scale bar, 200 µm. (E) Kaplan–Meier survival curves of mice implanted with BT025 cells transduced as in A–D. n = 7 mice for Rab35 nt63, n = 7 mice for ctrl. Statistical significance was examined by Mantel–Cox test (P = 0.0029).