Figure 1

Expression levels of class I and II HDACs in NSCLC cell lines and tumor tissue. (A) RNA was isolated from H23 and A549 cells, transcribed in cDNA and expression levels were analyzed by qRT-PCR. For normalization β-actin was used as a reference and data were presented as mean ΔCt values ± SEM. (B) RNA was isolated from fresh post-operative lung carcinomas. After reverse transcription, qRT-PCR was performed and expression levels of different HDACs in tumor tissue were normalized to β-actin and data were presented as mean ΔCt values ± SEM. (C, D) For protein analysis cryo-preserved lung tumor and non-malignant lung tissue was homogenized by blender, followed by sonication (3 x 5 seconds on ice) and centrifugation at 13000 rpm for 10 min. at 4°C in order to remove insoluble cell-debris. Tissue lysates (20 μg total protein) were analyzed by immunoblotting for expression of HDAC1 and HDAC4 in different tumor samples (C) and in comparison to corresponding non-malignant lung tissue (D). (C) Corresponding mRNA expression data are shown. ß-actin = loading control.