Our data highlight the possibility that because perturbations in pattern occur through delays early in the third instar, there is an intrinsic checkpoint late in the third instar that regulates pattern in the discs so that they reach a common patterning stage at pupariation

Our data highlight the possibility that because perturbations in pattern occur through delays early in the third instar, there is an intrinsic checkpoint late in the third instar that regulates pattern in the discs so that they reach a common patterning stage at pupariation. The progression of pattern also varied with genetic background. wing pouch along the dorsal-ventral axis. Stripes also appear as lines restricted to the anterior side of the wing pouch or along the dorsal-ventral boundary with a surrounding ring (stripe with ring, f), or along the anterior-posterior axis (g). The stripes correspond to lines of SOPs along the wing margin (e), or lines of positional information regarding wing disc boundaries (d). Different compositions of these elements describe all KPSH1 antibody observed patterns for each gene product through development.(TIF) pgen.1004408.s001.tif (368K) GUID:?69498767-044F-4B23-830F-4C4C3D37F256 Figure S2: Patterning progression of three of the eleven gene products initially assessed but not included in the staging scheme. Dynamic expression of Delta (1a-1j), Hindsight (2a-2j) and Scabrous (3a-3j) at 0 (1a-3a), 5 (1b-3b), 10 (1c-3c), 15 (1d-3d), 20 (1e-3e), 25 (1f-3f), 30 (1g-3g), 35 (1h-3h), and 40 (1i-3i) hours after third instar ecdysis (h AL3E) and wandering (1j-3j). Arrows show addition or change of cells or patches of cells, and asterisks highlight changes in stripes. (1f) Arrows highlight Delta expression mainly in the hinge and notum. Hindsight undergoes four transitions adding new elements at 15, 25 and 35 h AL3E. Lastly, Scabrous Sinomenine hydrochloride undergoes four transitions adding new elements at 10, 15 and 40 h AL3E. (3h) shows Scabrous expression in the centre of the wing pouch, before it refines to a stripe. (Scale bar 100 m). (k-o) Quantitative measures Sinomenine hydrochloride of the relative amount of expression normalized to disc size of the different elements observed. (k) Delta expression pattern represented by the progression of the stripe. (l-m) Hindsight expression pattern decomposed into (l) pattern area and (m) progression of the Sinomenine hydrochloride stripe. (n-o) Scabrous expression pattern decomposed into (n) pattern Sinomenine hydrochloride area and (o) progression of the stripe. Delta undergoes four transitions in its pattern, adding new elements at 5, 35 and 40 h AL3E.(TIF) pgen.1004408.s002.tif (1.6M) GUID:?4E4BEF5A-AEEC-439A-A0A8-8D3FA94D923A Figure S3: Quantitative measures of the relative amount of expression normalized to disc size and of the different elements observed for six of the eleven gene products. (a-c) Achaete expression pattern decomposed into (a) pattern area, (b) number of patches of cells and (c) progression of the stripe. (d-f) Cut pattern broken down into (d) pattern area (whole disc, only notum and hinge, and only wing pouch), (e) number of patches of cells and (f) progression of the stripe. (g) Notch expression pattern represented by the stripe progression. (h-k) Senseless expression pattern decomposed into (h) pattern area, (i) number of SOPs, (j) progression of the stripe and (k) number of doublets. (l-n) Dachshund expression pattern in terms of (l) pattern area, (m) number of patches of cells and (n) progression of the stripe. (o) Wingless pattern area in the whole disc, only notum and hinge, and only wing pouch.(TIF) pgen.1004408.s003.tif (871K) GUID:?8A4F7245-EDFA-4DB5-895E-F6DB4577EFC0 Figure S4: The progression of pattern, in absolute time, in discs from larvae with altered developmental time and from two parental lines. The probability (represented by the size of the circle) that a disc with a particular set of gene-specific stages belongs to a given disc stage, varied with absolute developmental time (hours after third instar ecdysis (h AL3E)). (a-b) Temperature manipulations include (a) 18C in blue and (b) 29C in orange. (c-d) We manipulated the timing of ecdysone synthesis using (c) larvae (in green) and (d) larvae (in purple). (e-f) Parental lines to test for the contribution of genetic background include (e) in red and.