(A) Scheme summarising the critical role of the brain hormone nereidin in energy expenditure, as derived from classical experiments: Before maturation (left), high nereidin levels sustain somatic …
Data for the graphs in Figure 1C, Figure 1—figure supplement 2A, Figure 1—figure supplement 2B (vtg expression levels over the time course of maturation).
Alignment file for the phylogenetic tree of Vitellogenins (Figure 1—figure supplement 1B).
(A) Domain analysis for the protein encoded by the identified Platynereis cDNA (top) as well as chicken Vtg2 (middle) and mosquito Vit1 (bottom) reveals that Platynereis Vtg shares three conserved …
The figure shows the same data as Figure 1C, but with the results for the individual reference genes plotted separately. In the range relevant for the bioassay (early premature stage), the observed …
(A–C) Nereidin activity elutes in lipophilic fractions of head extracts but is not proteinaceous as previously hypothesised. (A) Schematised fractionation of premature Platynereis head extracts, …
Data for Figure 2B,C,E,F,G (vtg expression levels normalised to the respective control treatment) and Figure 2—figure supplement 2 (stability of Ct values for rps9).
(A,B) The panels shows the selected ion currents (SIC) in the range of m/z 251.0–251.5 (top panels) and the selected MS2-spectrum of the precursor ion at m/z 251.1 (lower panels). (A) Peak 15. Note …
Ct values for rps9 were normalised to the mean of each experimental series. The box shows the first and third quartile and the median (solid line). Whiskers denote the 1.5x fold range of the …
(A) Significant decrease of MF levels between premature and mature heads; boxplots show amounts of MF normalised to the total protein content of the respective head sample. Heads of premature …
Data for the graphs in Figure 3A (MF content in pg normalised to mg protein per head), Figure 3C (met expression levels normalised to premature eleocytes) and Figure 3D (met expression levels normalised to the control treatment); data for the graphs in Figure 3—figure supplement 2 and Figure 3—figure supplement 3 (MF content in pg per head), Figure 3—figure supplement 4A and Figure 3—figure supplement 4B(met expression levels quantified against individual reference genes).
Alignment file for the phylogenetic tree of Met homologs (Figure 3B).
Representative Selected Ion Mode (SIM) chromatograms from premature Platynereis heads and authentic MF (upper panel). The lower panel shows the selected ions at m/z 114.0, 136.0 and 219.0. (A) SIM …
Significant decrease of MF levels between premature and mature heads; the boxplot shows the quantified amounts of MF normalised per head. Heads of premature animals contain on average ~ 34.6 ± 8.0 …
The head of earthworms contain on average ~ 5.3 ± 1.9 (S.E.M) pg MF per head. The data range is between ~2.1 and ~9.8 pg MF per head. The amounts are corrected for a MF recovery rate of 85%. The box …
The figure shows the same data as Figure 3C, but with the results for the individual reference genes plotted separately. The observed relative expression of met in eleocytes is independent of the …
(A) Setup of experiment testing the ability of MF to interfere with vitellogenesis in vivo. Following decapitation, female individuals are known to start vitellogenesis, reflecting loss of nereidin; …
Data for the graphs in Figure 4B and Figure 4C (vtg/hox3 expression levels normalised to the respective control treatment).
The figure shows the same data as Figure 4B, but with the results for the individual reference genes plotted separately. The observed relative expression of vtg in whole animals treated with either …
Expression levels of vtg in coelomocytes are significantly down-regulated by treatment with 10 nM methoprene (Meth) and 10 nM pyriproxyfen (Pyr) in 0.01% DMSO. The graph shows qRT-PCR quantification …
Data for the graph in Figure 5 (vtg expression levels normalised to the control treatment).
List of primers used for qRT-PCR, including predicted melting temperature.
Name | Sequence 5’→3’ | Tm / °C | Efficiency |
---|---|---|---|
Pdu vtg qPCR1 F | ACAGGCCATCACATTCACAA | 56.4 | 101% |
Pdu vtg qPCR1 R | TCTGCTCACGTCTCTTTCCA | 58.4 | |
Pdu met qPCR1 F | GGATGATTATGATGTATACCTGCAAC | 62.9 | 102% |
Pdu met qPCR1 R | AGACCGAACTGGCGTTTG | 56.3 | |
Pdu hox3 qPCR1 F | CTACCCCTGGATGAGGGAAT | 60.5 | 95% |
Pdu hox3 qPCR1 R | ACTTCCGGTTCCTGGTCC | 58.4 | |
Pdu rps9 F | CGCCAGAGAGTTGCTGACT | 59.5 | 102% |
Pdu rps9 R | ACTCCAATACGGACCAGACG | 60.5 | |
Pdu sams qPCR1 F | CAGCAACGGTGAAATAACCA | 56.4 | 101% |
Pdu sams qPCR1 R | CATCACTCACTTGATCGCAAA | 57.5 | |
Pdu cim6pr qPCR1 F | ACTTCCCCTGCTGATGAGTG | 60.5 | 99% |
Pdu cim6pr qPCR1 R | TTCGTAAGTCAGGTTTCCATTG | 58.4 | |
Pdu cdc5 F | CCTATTGACATGGACGAAGATG | 60.1 | 100% |
Pdu cdc5 R | TTCCCTGTGTGTTCGCAAG | 57.5 |
List of primers used for cloning and sequence validation.
Name | Sequence 5’→3’ | Tm / °c |
---|---|---|
Pdu vtg 1F | ATGAAGACTCTCCTGATCTTCG | 60.1 |
Pdu vtg 1R | CTAGTAGTAGAATCTTGGTCCTTCAC | 64.6 |
Pdu vtg_seq 1F | AGCCCTAGAAGCTGCCTCTG | 62.5 |
Pdu vtg_seq 2F | ATTGCTCAATCTGAACTCCCATGC | 63.6 |
Pdu vtg_seq 3F | GCTGTTCCACAGGAAATTGC | 58.4 |
Pdu vtg_seq 4F | GCTTTGGTCAGTGGACTTCC | 60.5 |
Pdu vtg_seq 1R | GGCAATCCTCTGATGTAAACATTCTC | 64.6 |
Pdu vtg_seq 2R | CAAGCGTTTCACGACCAAGAGG | 64.2 |
Pdu vtg_seq 3R | GAAGAGCTTCTTGCTGGAGC | 60.5 |
Pdu vtg_seq 4R | AAGACCAGCTGGCGCGTTATG | 63.2 |
Pdu met F | ATGGAGCCGAATTCGGAGCAGAATTCGG | 71.8 |
Pdu met R | TCAACATGTCTCAGTTTCTTTTTGAGCG | 65.6 |
Pdu hox3 F | CCCCGGGGCTCTTGGTTTT | 61.6 |
Pdu hox3 R | GCCATCTCTATTCTCCTCGGCCG | 68.3 |