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Files in this Data Supplement:
Fig. S1. Detection of different Drosophila TFIIH components by western blot and identification of XPB and Cdk7 in cytoplasmic and nuclear fractions from cellularized embryos. (a) Protein extracts from 0-20 hour embryos were loaded in SDS-PAGE gels and after electrophoresis transferred to a nitrocellulose filter. The filters were then incubated with the corresponding antibody. The antibodies used were against XPB, XPD, Cdk7 MAT1 proteins, each one indicated in the figure. (b) N represents the nuclear fraction and C1-3 lanes are different cytoplasmic fractions. The same blot was treated with both antibodies (anti-XPB and anti-Cdk7) simultaneously. Note that Cdk7 is preferentially located in the cytoplasm and DmXPB in the nuclear fraction. (c) Detection of the SOD enzyme in different cellular fractions by western blot. SOD was used as control to confirm the absence of cross contamination between the cytoplasmic and nuclear fractions. The antibodies used in this control experiment are indicated in the figure. T.E., total cellular sample; Nuc., nuclear fraction after enrichment by differential centrifugation with the transcriptionally active chromatin; Cit., cytoplasmic fraction. Note that SOD is absent in the nuclear fraction.
Fig. S2. XPD distribution in early fly development and co-localization with XPB (haywire). (a-d) Staining pattern of DmXPD (shown in red) from early syncytial blastoderm stage (a), to cellularization (d). DNA is in green. (e) Amplification of a stage 10 embryo showing nuclei in the embryo periphery that contain XPD. (f) Amplification of the signal from the cellularized embryo. Most of the XPD signal is in the nucleus. (g) Section of a gastrulated embryo. Note that chromosomes in mitosis do not have XPD (arrow), but the amount in the cytoplasm is not reduced. This pattern is identical to that observed with XPB (Fig. 1). (h-j) Co localization of XPB (red) and XPD (green) in early fly embryogenesis.
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