Fig. 4. smt3i PG cells show changes in steroidogenic enzymes and transcription factors. (A-O) Single plane confocal micrographs showing expression of the indicated hydroxylase enzymes (Phm, Dib or Sad) or the indicated transcription factors (Woc, Mld or βFtz-f1) in WT, smt3i or lwr mutant PG cells. Nuclei are labelled with DAPI and shown in purple; the indicated proteins are shown in green. (A'-O') Single green channels for each panel, showing expression of the indicated proteins, are presented in black and white. White arrows denote differences in the nuclear accumulation of the referred factors between WT and knockdown phenotypes, and white arrowheads denote ectopic cytoplasmic accumulation of certain factors in knockdown or mutant backgrounds. (A,A',D,D') Phm, accumulated in the ER, does not show changes in expression levels or localisation in smt3i larvae compared with WT, whereas Dib mitochondrial staining is reduced (B,B',E,E'). Sad nuclear accumulation, but not mitochondrial staining, is reduced in knockdown larvae (C,C',F,F', arrows). (G,J,M) Woc nuclear localisation does not diminish in smt3i (J,J') or lwr mutant larvae (M,M') compared with WT (G,G'). In lwr mutants, Woc accumulates in the cytoplasm (arrowhead). (H,K,N) Mld accumulates in the nucleus in WT cells (H,H', arrow), and is highly reduced in smt3i nuclei (K,K', arrow), although cytoplasmic accumulations of the protein can be observed (arrowheads). This is even more noticeable in lwr mutant larvae (N,N', arrowheads). (I,L,O) βFtz-f1 appears to be evenly localised in the nucleus (arrow) and the cytoplasm in WT cells (I,I'), whereas in smt3i (L,L') and lwr mutant larvae (O,O') it is reduced in the cytoplasm and absent in the nuclei (arrows). All images are shown at the same magnification.