Figure 8.

BTN3A1 expression in CRC tissue specimens. BTN3A1 expression was evaluated in CRC tissue sections by Q-RT-PCR (A), western blot (B) or immunohistochemistry (C). (A) RNA was extracted from tissue sections of 10 CRC specimens, reverse transcribed and Q-RT-PCR for BTN3A1 performed. Results are expressed as1/ΔCt normalized to 18s. (B) immunoblots of lysates obtained from the indicated CRC tissue specimens, as described in Materials and Methods, with the anti-CD277 mAb (upper blot) or with a rabbit polyclonal anti-BTN3A1 antiserum (lower blot). β-actin is shown as a loading control. (C) immunohistochemistry of two representative cases out of the 10 indicated in panels A and B, performed as described in Materials and Methods, with the indicated antibodies: polyclonal rabbit anti-BTN3A1 antiserum (arrows), anti-Vδ2 mAb (BB3, arrows in the inset), polyclonal rabbit anti-TGII antiserum, anti-vimentin mAb and a matched isotype-unrelated antibody as negative control (goat anti-rabbit antiserum in the inset of the upper CTR). Slides were counterstained with hematoxylin, coverslipped with Eukitt and analyzed under a Leica DM MB2 microscope with a charged-coupled device camera (Olympus DP70) at a 40× enlargement, as indicated.