Figure 1

Hdac1 depletion alters IEC-6 cell proliferation and deacetylase activity. A. Total RNAs were extracted from two shCtrl (5; 9) or shHDAC1 (18; 21) IEC-6 cell lines. Class I Hdac1, Hdac2, Hdac3 and Hdac8 mRNA levels were measured by semi-quantitative RT-PCR, with Gapdh as a control for the amount of cDNA amplified. Results are representative of two independent experiments. B. Nuclear protein extracts from shCtrl (5; 9) and shHDAC1 (18; 21) IEC-6 cells were subjected to Western blot analysis, with Hdac1 and Hdac2 specific antibodies. Lamin B (Lmnb1) detection was used as a control for protein loading. C. 10,000 shCtrl (5; 9) and shHDAC1 (18; 21) IEC-6 cells were plated in each well of a 24-well plate. Cell growth was measured by cell counting with a hemocytometer between 3 and 8 days after seeding. Results are representative of two independent experiments. Differences in cell numbers between shCtrl and shHDAC1 cells are statistically significant (^* p ≤ 0.05; ^** p ≤ 0.01). ▲, shCtrl 5; ■, shCtrl 9; ♦, shHDAC1 18; ●, shHDAC1 21. D. Nuclear proteins from shCtrl and shHDAC1 IEC-6 cells were immunoprecipitated with specific antibodies against Hdac1, Hdac2 and mSin3a. Deacetylase activity was measured using a commercial colorimetric HDAC assay kit. Deacetylase activity was calculated in pmoles/min/mg. Results are representative of two independent experiments.