Frames were acquired for 24 moments in one-minute intervals using a spinning disc confocal microscope (CSU-10; Yokogawa Electric Corporation). AU = arbitrary devices; scale pub, 5 m.(TIF) pgen.1005786.s005.tif (1.7M) GUID:?45446CED-921C-4ACF-860B-CB0460B528B8 S3 Fig: is required for proper trafficking of the invadopodial membrane. (A-E) 3D renderings showing the distribution of PI(4,5)P2 (A, (middle panels) COTI-2 resulted in mis-trafficking of the invadopodial membrane parts PI(4,5)2, GFP::MIG-2, and GFP::CED-10, as well as GFP::CUP-5 and LMP-1::GFP (which are found both MAD-3 in the invadopodial membrane and the endolysosome) relative to crazy type (remaining panels). RNAi focusing on of did not impact COTI-2 the distribution of PI(4,5)P2, GFP::MIG-2, or GFP::CUP-5 (ideal panels). Package plots (collection shows median, boxes cover the interquartile range, and bars show minimum and maximum) display the percentage of the total fluorescent transmission at or near the basal invasive cell membrane of the AC. For those conditions a minimum of 9 animals were analyzed (n is definitely mentioned on each graph). In (A-C) comparisons were made using Tukeys multiple comparisons checks, ** p < 0.01, *** p < 0.001. In (D-E) comparisons were made using a College students t-test, * p < 0.05. Level pub, 5 m.(TIF) pgen.1005786.s006.tif (540K) GUID:?FA1EB4EE-0755-4069-B87B-6451257ADFC7 S1 Movie: Wild type AC invadopodia dynamics. Ventral look at time-lapse showing spot tracking analysis of a crazy type animal prior to BM breach. Invadopodia are designated by F-actin (RNAi treated animal prior to BM breach. Invadopodia are designated by F-actin (reduced the number invadopodia and decreased the pace of invadopodia formation but did not affect invadopodia lifetimes. Frames were acquired for 24 moments in one-minute intervals using a spinning disc confocal microscope (CSU-10; Yokogawa Electric Corporation). The frames are played at 10 frames per second. This video corresponds to Fig 1E and 1F and Table 2. Scale pub, 5 m.(MOV) pgen.1005786.s008.mov (3.7M) GUID:?B59FF0B6-E924-4869-9E59-3415A4028F33 S3 Movie: AC invadopodia dynamics after loss of the vulval precursor cells. Ventral look at time-lapse showing an AC of a RNAi treated animal prior to BM breach. Invadopodia are designated by F-actin (RNAi) and thus loss of a cue(s) generated from the vulval precursor cells reduced the number invadopodia, decreased the pace of invadopodia formation, and improved invadopodia lifetimes. Frames were acquired for 24 moments in one-minute intervals using a spinning disc confocal microscope (CSU-10; Yokogawa Electric Corporation). The frames are played at 10 COTI-2 frames per second. This video corresponds to Fig 3D and 3E and Table 2. Scale pub, 5 m.(MOV) pgen.1005786.s009.mov (3.7M) GUID:?71F7009F-0C77-4F5B-ACF7-5AC82D25BBE7 S4 Movie: AC invadopodia dynamics after loss of RNAi treated animal prior to BM breach. Invadopodia are designated by F-actin (by RNAi reduced the number invadopodia, decreased the pace of invadopodia formation, and improved invadopodia lifetimes. Frames were acquired for 24 moments in one-minute intervals using a spinning disc confocal microscope (CSU-10; Yokogawa Electric Corporation). The frames are played at 10 frames per second. This video corresponds to Fig 4C and 4E and Table 2. Scale pub, 5 m.(MOV) pgen.1005786.s010.mov (3.7M) GUID:?69DA591E-6E60-4983-BA57-B7227CF39866 S5 Movie: The AC invadopodial membrane dynamically traffics to the invasive cell membrane. A 3D reconstruction of a lateral look at time-lapse of a crazy type AC visualizing the invadopodia membrane having a probe for PI(4,5)P2 (cyan; RNAi treated AC visualizing the invadopodial membrane component PI(4,5)P2 (cyan; resulted in mis-trafficking of the invadopodial membrane to lateral and apical plasma membrane. The time-lapse takes place over a 40-minute period. The images were acquired in one-minute intervals using a spinning disc confocal microscope (CSU-10; Yokogawa Electric Corporation). The frames are played at 10 frames per second. This video corresponds to Fig 6B. Level pub, 5 m.(MOV) pgen.1005786.s012.mov (5.5M) GUID:?FDFA367D-A407-41AE-AD37-69A5B15EE72E S7 Movie: The invadopodial membrane is definitely mis-trafficked after loss of the vulval precursor cells. A 3D reconstruction of a lateral look at time-lapse of a RNAi treated AC visualizing the invadopodial membrane component PI(4,5)P2 (cyan; RNAi treatment results in loss of the vulval precursor cells, which inhibits generation of the vulval cue. Loss of the vulval precursor cells resulted in mis-trafficking of the invadopodial membrane to the lateral and apical plasma membrane. The time-lapse takes place over a 40-minute period. The images were acquired in one-minute intervals using a spinning disc confocal microscope (CSU-10; Yokogawa Electric Corporation). The frames are played at 10 frames.