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. 2009 Oct 19;119(11):3408–3419. doi: 10.1172/JCI38854

Figure 5. Fanc/Brca pathway gene expression and function correlate with Smad4 expression levels.

Figure 5

(A) Relative expression levels of Fanc/Brca transcripts examined by qRT-PCR after Smad4 restoration in Smad4-deficient HNSCC cells (Cal27). Brca1 and Rad51 expression was increased by Smad4 restoration in Cal27 cells. The expression level of each gene in parental Cal27 was arbitrarily set at 100% for each experiment. Samples were run in triplicate for each experiment, and the mean relative expression levels from 3–4 independent experiments are presented. Error bars indicate SEM. *P < 0.05 versus parental Cal27 cells. (B) Relative expression levels of Fanc/Brca transcripts examined by qRT-PCR after Smad4 knockdown in normal keratinocytes (HEKn). Brca1, FancA, FancD2, and Rad51 expression was decreased after 48 hours of Smad4 siRNA treatment. The expression level of each gene in untreated HEKn cells was arbitrarily set at 100% for every experiment. Samples were run in triplicate for each experiment, and the mean relative expression levels from 3–4 independent experiments are presented. Error bars indicate SEM. *P < 0.05 versus untreated HEKn cells. (C) Brca1 and Rad51 nuclear foci detected by immunofluorescence in Cal27 and Cal27-Smad4 cells after MMC treatment. A representative image is presented. The histogram indicates the percentage of cells with Brca1 or Rad51 foci in Cal27 and Cal27-Smad4 cells. One hundred to 200 cells per group were analyzed. Error bars indicate SEM. *P < 0.05 versus Cal27. Scale bar: 5 μm (all panels).