JCS01113 Supplemental Figure

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• Figure S1 -

  Fig. S1. FHOS is differentially expressed in human tissues and cell lines. (a) An ~4 kb message was detected in total RNA prepared from the following human tissues: heart, brain, kidney, liver, lung, pancreas, spleen, and skeletal muscle. RNA blot hybridization was performed using a radiolabeled C-terminal fragment of FHOS (see below). GAPDH expression was concurrently analyzed as a control for RNA loading. (b) Analysis of the distribution of FHOS in mature and fetal tissues was performed using a human MTE Array blot (Invitrogen) probed with the radiolabeled C-terminal fragment of FHOS (see below).

  RNA blot hybridization

  The C terminus of FHOS was amplified using specific primers (MG112 and MG113, as previously detailed) and radiolabeled using the Random Primed Labeling kit (Invitrogen). A master blot filter containing 20 mg of total RNA from eight human tissues (Invitrogen) was pre-hybridized overnight at 42°C and then re-hybridized overnight at 42°C with a radiolabeled FHOS probe (1.5´10⁹ cpm/mg). Filters were washed 4´ at room temperature in 2´ SSC and 0.01% SDS for 15 minutes and once at 68°C in 0.2´ SSC/0.05% SDS for 1 hour. Autoradiography was performed using X-Omat film (Kodak) with intensifying screens at –70°C. A human Multiple Tissue Expression Array (Clontech) was hybridized with the identical probe.