Figure 1.

Analysis of the glucose transporters GLUT1-3 in the primary uveal melanomas (UMs) by immunohistochemistry. (A) Representative images from the immunostainings for GLUT1-3. Signal detection was performed using the horseradish peroxidase-green substrate, which generates a blue-green reaction product. The nuclei were counterstained with nuclear fast red. The strength of the immunohistochemical reaction within a circumscribed area was graded using a scale of 0–3 (0: negative. 1: weak. 2: intermediate. 3: strong). The mean intensity of the entire tumor area was determined by a semiquantitative approach as described in the Methods section. Images were acquired at an original magnification of 200×. Arrows indicate the pigmented regions. Scale bar = 25 μm. (B) Immunoblots with GLUT1-3 peptides or full-length recombinant human GLUT3 protein (f), demonstrating the specificity of the antibodies used. The total protein amount in the wells was detected by the stain-free imaging of gels after ultraviolet-activation. The white bands (arrows) correspond to the bromophenol blue dye in the loading buffer, which has not migrated out of the gels. kDa: Kilodalton. (C) Quantification of the GLUT1, GLUT2, and GLUT3 intensities in the primary UMs with regard to the monosomy-3 status. The sum of the GLUT1-3 intensities was also presented as “total”. p-values were determined by the Mann-Whitney U test. (D) Ratios of GLUT1, GLUT2, and GLUT3 to the total GLUT1-3 levels in the primary UMs with respect to the monosomy-3 status. p-values were analyzed by the Mann-Whitney U test. The significant p-values (<0.05) in (C,D) were stated in red.