Figure 4. Effects of insertions and deletions of the class I and class II dKdm2 alleles on the expression of dKdm2 and its neighboring genes.

(A) The location of the primers used for the qRT-PCR assay. Animals of the third instar at the wandering stage were used for the qRT-PCR (C ~ F) and immunoblots (G and H). The genotypes of the mutants include: w¹¹¹⁸; +; Df(3R)J15 (B, first instar laevar), w¹¹¹⁸; +; Df(3R)J16 (C), w¹¹¹⁸; +; Df(3R)J18 (D), w¹¹¹⁸; +; dKdm2^d00170 (E), and w¹¹¹⁸; +; dKdm2^DG12810 (F). (G) Western blot was used to demonstrate the specificity of the dKDM2 antiserum, note that the ~150kDa band is present in the control (w¹¹¹⁸) but not in the Df(3R)J16 and dKdm2^DG12810 mutants. In addition, this band is present in untreated S2-DRSC cells or cells treated with dsRNA to white (control), but disappears in S2-DRSC cells treated two different shRNAs targeting dKdm2. (H) Western blot of nuclear soluble proteins from the third instar larvae, showing that dDKM2 protein is undetectable in Df(3R)J16, Df(3R)J18, dKdm2^d00170 or dKdm2^DG12810 homozygous mutants.