Figure 4. A. Detection of polyP associated with the TRPM8 protein. TRPM8 was separated on native PAGE to preserve its migration in the tetrameric form. Lane 1 – standards ladder (The High-Mark Pre-stained High Molecular Weight Protein Standards, Invitrogen); Lane 2 – purified TRPM8 sample with o-toluidine blue stainof native PAGE gel; Lane 3 – o-toluidine blue stain of native PAGE gel of the same TRPM8 sample treated with 1 μl scPPX1 (2 μg/ml) for 3 h. before loading: Lane 4 and 5 are lanes 2 and 3 re-stained with Coomassie blue. B. Detection of PHB in TRPM8 in Western blot. Lane 1 – purified TRPM8 protein detected with antiMyc_IgG; Lane 2 – Western blot of purified TRPM8 probed with anti-PHB-IgG. Samples were heated for 5 min. at 70 °C before loading.