Western blotting was conducted using rabbit polyclonal anti-GilZ/TilZ antibody (1 1000) (Abcam) (Tse et al., 2012), anti-phospho-Akt, anti-total Akt, anti-phospho-GSK3β or anti-total GSK3β (1:1000) (Cell signaling), followed by an incubation with horseradish peroxidase-conjugated goat anti-rabbit antibody (1 4000) (Bio-Rad).
ChIP assays and real-time quantitative (q PCR experiments to determine Spt16 occupancy at the different regions tested in this study were conducted using rabbit polyclonal antibodies specific for Spt16 (a gift from Tim Formosa) as previously described (Myers et al. 2011).
Localization studies were conducted using affinity-purified polyclonal mouse anti-TbUGP antibody together with either rabbit anti-GAPDH antibody (as a glycosomal marker) or rabbit anti-enolase (as a cytosolic marker).
To confirm surface-exposure of these proteins in transformed B. garinii, immunofluorescence microscopy was conducted using live bacteria and polyclonal antibodies specific for either CRASP-3 or CRASP-5 [40].
Immunoblot experiments were conducted using rabbit anti-gfp polyclonal antibody (gift from Dr. B. Moffatt, Waterloo).
Immunoblotting was conducted using anti-EZH2 rabbit polyclonal antibody (D2C9, 1 : 1000 dilution; Cell Signaling, Beverly, MA, USA) overnight at 4 °C.
IHC analysis of TRPM7 expression was conducted using goat anti-human TRPM7 polyclonal IgG antibodies (Abcam®, Cambridge, Massachusetts, USA) at a 1 50 dilution.
These experiments were conducted using 1 μL of the following antibodies: rabbit polyclonal antibody specific for the Spt16 protein (a generous gift of Tim Formosa), a mouse monoclonal antibody specific to the yeast Pol II subunit Rpb3 (Neoclone, no. W0012), and a rabbit polyclonal antibody specific for bulk histone H3 (ab1791; Abcam).
Immunostains were conducted using mouse monoclonal anti-tyrosine hydroxylase (anti-TH) antibody (Millipore, Billerica, MA), rat polyclonal anti-DAT (Millipore, Billerica, MA), rabbit polyclonal anti-GFAP (Millipore), and rabbit polyclonal cleaved-caspase-3 (Cell Signaling Technology, Beverly, MA) with antigen-matched blocking peptide (Cell Signaling Technology).
In 41 out of 55 carcinomas, immunohistochemistry was conducted, using monoclonal antibodies against bcl-2, fas and DNA fragmentation, and polyclonal antibody for bax.
