A complex containing the CCR4 and CAF1 proteins is involved in mRNA deadenylation in Drosophila - PubMed (original) (raw)

Figure 5

Expression and subcellular distribution of CCR4 and CAF1 in Drosophila. (A, B) Western blots probed with anti-CCR4 (A) and affinity-purified anti-CAF1 (B) showing the presence of CCR4 and CAF1 in different developmental stages, and the lack of CCR4 in ccr4 mutants (A, right panels). Protein extracts loaded were from 0.75 ovary, 20 embryos, 20 first instar larvae, 0.75 third instar larva and 0.3 adult male. Shown are wild-type extracts (A, lanes 1–9, 12 and B, lanes 1–8), extracts from ccr4 KG877/ccr4 KG877 first instar larvae, taken 5–15 h after eclosion (A, lane 10, y: young) or about 30 h after eclosion (A, lane 11, o: old), and extract from ccr4 KG877/Df(3R)crb-F89-4 adult males (A, lane 13). The blots were then probed with anti-α-tubulin as a loading control. (C–P) Immunodetection of CCR4 and CAF1 in ovaries. (C, D, J, K) Confocal images showing that CCR4 and CAF1 are present at all stages of oogenesis and in the different cell types. Earliest stages of oogenesis (left panels), stage 10 (right panels), anti-CCR4 (C), anti-CAF1 (J) and DAPI (D, K). (E–I, L–P) Enlargement of stage-10 egg chamber follicle cells showing accumulation of CCR4 (E–G) and CAF1 (L–N) in cytoplasmic foci in the wild type and a strong decrease in CCR4 (H, I) and CAF1 (O, P) levels in ccr4 KG877/Df(3R)crb-F89-4 mutant females. Anti-CCR4 in red (E, G, H, I), Anti-CAF1 in red (L, N, O, P), DAPI in blue (F, G, I, M, N, P), merge anti-CCR4/DAPI (G, I) and anti-CAF1/DAPI (N, P).