Supplemental Figure 1
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Fig. S1.
Site-specificity of anti-phospho-H3 antibodies by peptide competition. Quiescent C3H 10Tg cells were
treated with 25 ng/ml anisomycin (lanes 2, 4, 6, 8) or left untreated (lanes 1,
3 5, 7). Acid-extracted proteins (as described in Material and Methods) were
run on 15% SDS-PAGE and Coomassie-stained to show equal protein loading (panel
x) or transferred to PVDF-membrane for western blotting with anti
phospho-S10occ. (panels i-iii), anti-phospho-S10 (panels iv-vi) or
anti-phosphoS28 (panels vii-ix). For competition analyses, antibodies were
pre-incubated with 10 mg/ml phosphoS10
peptide (panels ii, v, viii) or phosphoS28 peptide (panels iii, vi, ix). These
analyses show that the relevant phosphopeptides block the corresponding
antibody with high specificity and there is no cross-specificity of these
antibodies against the non-cognate phosphopeptide.
