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Identification of Genes Differentially Expressed in Cultured Human Periodontal Ligament Fibroblasts vs. Human Gingival Fibroblasts by DNA Microarray Analysis

Department of Periodontology & Oral Biology, Goldman School of Dental Medicine, Boston University, 100 East Newton Street, G05, Boston, MA, 02118, USA;

View larger version (94K): [in a new window]   Figure 1. Expression of periostin, S100A4, and CD40 in PDLF and GF analyzed by Northern blot. A pool of total RNA (20 µg) was isolated from cultured PDLF or GF from three donors. Equal amounts of RNAs were used from each cell passage. RNAs were blotted and hybridized serially with each cDNA probe for periostin, S100A4, and CD40. Blot was finally hybridized with a probe for human glyceraldehydes-3-phosphate dehydrogenase (GAPDH) as an internal standard. The results were representative of three independent repeats (n = 3).

View larger version (63K): [in a new window]   Figure 2. Differential expression between PDLF and GF of the 9018 unique human cDNAs represented in the microarray. mRNA from PDLF was labeled with Cy5; mRNA from GF was labeled with Cy3. Forward gridlines indicate expression ratios between the two probes. Each dot represents a gene. Genes differentially expressed by at least three-fold were indicated as those above or below red dashed lines.

View larger version (92K): [in a new window]   Figure 3. Confirmation of differentially expressed genes observed in microarray results. (A) Four genes (MFG-E8, MRPL3, TMP2, and STK) selected from array results were analyzed by Northern blot as described in Fig. 1. (B) Radioactive signals were quantified by PhosphorImager with IMAGEQUANT software (Molecular Dynamics). Human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA probe was used as an internal control. Data are presented as mean ± standard deviation and expressed as ratios relative to GAPDH values (*p < 0.05, **p < 0.01, n = 3).