Figure 5

The Cp190 fragment lacking the C-terminal E-rich domain localizes to Cp190-containing nuclear complexes. (A) Polytene chromosomes from y²; CP190^En15/CP190^P11(top and middle panels) and from y²; CP190^En15mod(mdg4)^T6/CP190^P11mod(mdg4)^T6(bottom panel) 3^rd instar larvae were stained with anti-Su(Hw) (green, left column) and anti-Cp190 (red, middle column) antibodies. Merged images are shown on the right. The middle panel is the closer view, rotated 90 degrees counter clockwise, of the area indicated by the yellow square in the top image. White arrows point to the y locus at the tip of the X chromosome. Yellow arrows point to the Cp190-positive band at the constriction of the cytological location 3C. (B) Diploid cells of brain tissues from CP190⁺flies (top) or from the CP190^En15/CP190^P11(bottom) flies were stained with anti-Cp190 (green, left column) and anti-Mod(mdg4) (red, middle column) antibodies. Merged images are shown on the right. (C) The gypsy fragment amplified by PCR from anti-Cp190 precipitated chromatin from y²; CP190^En15/CP190^P11pupae (left lanes, labeled CP190^En15) and from y²; CP190⁺pupae (middle lanes). The gypsy fragment amplified from the pre-immune serum precipitated y²pupae (right lanes, labeled CP190⁺and Pre-Imm) was the negative control and the gypsy fragments amplified from non-precipitated total y²pupal genomic DNA (labeled as input on top) was the positive control. The black triangles indicate two-fold serial dilutions of the amount of the chromatin sample used in each PCR reaction. More volumes of the indicated template were in the PCR reactions on the left as indicated by the thicker part of each triangle. (D) The anti-Cp190 ChIP of y²ct⁶; CP190^En15flies analyzed by Real-Time PCR. The same loci tested in Figure 3 ChIP assays were analyzed and were shown as percentage of input DNA (n ≥ 3). The results of all regions were normalized to Fab-8.