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Figure 3 | BMC Medical Genetics

Figure 3

From: Cell type-specific over-expression of chromosome 21 genes in fibroblasts and fetal hearts with trisomy 21

Figure 3

Validation of p78 MX1 over-expression in +21 fibroblasts by western blotting, correlation of p78 MX1 protein with MX1 mRNA, and induction of MX1 mRNA in normal fibroblasts by interferon-beta present in +21 fibroblast-conditioned medium. A, p78MX1 is over-expressed in +21 fibroblasts, directly correlating with MX1 RNA. The top two panels show results from a western blot of proteins from whole-cell lysates of +21 (n = 4) and control (n = 3) fibroblasts at late passage. The blot was re-probed with anti-beta-actin is as a loading control. The bottom panel shows the relative MX1 RNA levels of all the samples from the GeneChip data, normalized to the value for the late-passage C1 fibroblasts. P, passage number. The excess of MX1 mRNA in these cells leads to the expected excess production of p78MX1 protein. B, conditioned medium from fibroblasts with trisomy 21 was added to the medium of control fibroblasts at the indicated proportions, either with our without the addition of anti-interferon beta antibodies. RNA was extracted after 24 hours and analyzed by northern blotting. Addition of anti-interferon almost completely abrogates the induction of MX1 mRNA by the +21 conditioned medium.

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