No adult flies after expression of the indicated dsRNA.(XLSX) pgen.1008253.s008.xlsx (33K) GUID:?D099B458-9027-4192-B2FB-59AA4BDAFE33 S8 Table: Full list of fly strains and primers used. pgen.1008253.s005.xlsx (21K) GUID:?25BF0A3A-7A5C-4D4A-98F0-F54550EC03D8 S5 Table: Differentially expressed (DE) genes shared by CycA dsRNA iECs, Myb dsRNA iECs and salivary gland devECs. (XLSX) pgen.1008253.s006.xlsx (67K) GUID:?9121B8F7-45DF-4516-88EB-8721A7392C72 S6 Table: Meta-analysis of RNA-Seq data for E2F1 regulated genes (from Dimova et al. 2003)(3). (XLSX) pgen.1008253.s007.xlsx (40K) GUID:?262E3071-EFAE-4D0A-9D0C-0EA5B467AB0A S7 Table: Results of RNAi wing screen. Stock#Bloomington Drosophila Stock Center (BDSC) Stock number. Light bluepositive hit in the wing screen. Reduced size of L3-L4 region, increased bristle size. OrangeLethal. No adult flies after expression of the indicated dsRNA.(XLSX) pgen.1008253.s008.xlsx (33K) GUID:?D099B458-9027-4192-B2FB-59AA4BDAFE33 S8 Table: Full list of travel strains and primers used. Stock#CBloomington Drosophila Stock Center (BDSC) Stock number.(XLSX) pgen.1008253.s009.xlsx (100K) GUID:?BF9FD4A7-5372-4073-B90C-1E11CCC74B1A S1 Fig: Knockdown of CycB does not induce endoreplication. S2 cells were treated with CycB dsRNA. (A) qRT-PCR quantification of CycB transcript in CycB dsRNA versus GFP dsRNA control cells. (B) Quantification of circulation cytometry data for ploidy classes in GFP dsRNA and CycB dsRNA cells (mean and S.D. for N Acetoacetic acid sodium salt = 2).(TIF) pgen.1008253.s010.tif (1.0M) GUID:?5B59AE13-341A-4F4E-8326-8CBE790C3986 S2 Fig: Knockdown of CycA or Myb inhibits cell proliferation. 500,000 cells were plated and treated with the indicated dsRNAs. The cells were counted once every 24h for 7 days (mean and S.D. for N = 3).(TIF) pgen.1008253.s011.tif (891K) GUID:?3DB59D89-946A-437A-8B2E-C4346C784FD1 S3 Fig: Statistical analysis of DE gene overlap between populations of endoreplicating cells. Permutation screening was used to determine animal. (B) A wing disc from a animal. Note the larger nuclei within the reddish border compared to cells outside. (C-I) Wing discs after expression of (C), (D), (E), (F), (H), or (I). Level bars are 20M.(TIF) pgen.1008253.s016.tif (5.5M) GUID:?7A815314-B579-4F46-9F4B-1234F08885DB S8 Fig: RT-qPCR quantification of RNAi knockdown in larval discs. RT-qPCR quantification of the indicated transcripts in imaginal discs from different UAS-dsRNA strains normalized to that in wild Acetoacetic acid sodium salt type control discs. Each value around the X axis indicates both the dsRNA strain and the transcript measured after induction with a warmth inducible GAL (N = 2).(TIF) pgen.1008253.s017.tif (1.2M) GUID:?5710FB7D-3F8E-457F-9894-66884B874893 S9 Fig: Knockdown of aurB induces endoreplication whereas knockdown of polo induces a mitotic arrest in S2 cells. (A) Circulation cytometry of DNA content Rabbit polyclonal to PSMC3 in propidium iodide labeled S2 cells 96 hours after treatment with either GFP dsRNA (control), aurB dsRNA or polo dsRNA. (B) Quantification of EdU and pH3 labeling in cells after treatment with the indicated dsRNAs (mean and S.E.M. for N = 3, *p < 0.05, ** p < 0.01, nsnot significant).(TIF) pgen.1008253.s018.tif (1.0M) GUID:?837F0A92-15B1-48C0-9FF8-ED2F5380DD2E S10 Fig: Myb over-expression does not inhibit endoreplication after CycA knockdown. Induction of endoreplication by knockdown of CycA is not suppressed by overexpressing Myb. Quantification of nuclear area of ovary follicle cells in stage 6 egg chambers after warmth inducing the following genotypes: 1) to determine how mitotic cycles are remodeled into endoreplication cycles, and how similar this remodeling is usually between induced and developmental endoreplicating cells (iECs and devECs). Our evidence suggests that Cyclin A / CDK directly activates the Myb-MuvB (MMB) complex to induce transcription of a battery of genes required for mitosis, and that repression of CDK activity dampens this MMB mitotic transcriptome to promote endoreplication in both iECs and devECs. iECs and devECs differed, however, in that devECs experienced reduced expression of E2F1-dependent genes that function in S phase, whereas repression of the MMB transcriptome in iECs was sufficient to induce endoreplication without Acetoacetic acid sodium salt a reduction in S phase gene expression. Among the MMB regulated genes, knockdown of AurB protein and other subunits of the chromosomal passenger complex (CPC) induced endoreplication, as did knockdown of CPC-regulated cytokinetic, but not kinetochore, proteins. Together, our results indicate that this status of a CycAMyb-MuvBAurB network determines the decision to commit to mitosis or switch to endoreplication in both iECs and devECs, and suggest that regulation of different actions of Acetoacetic acid sodium salt this network may explain the known diversity of polyploid cycle types in development and.