Supplementary Movies 1 and 2 show the entire time-lapse recording. confocal laser-scanning microscope (LSM 510; zoom factor 63X) to monitor PI entry. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s3.mov (1.4M) GUID:?A3824B9F-5B28-45E7-B816-DD6BD5319D2F Supplementary Movie 3 JY cells were seeded in chambered wells in the presence of 8g ml-1 Av-SRho (pseudo color) and monitored for 2 hours using a confocal spinning disk microscope to visualize constitutive vesicular exocytosis. A typical cell is shown; data are representative of three impartial experiments. ncomms10823-s4.mov (518K) GUID:?C366B203-5F9E-4150-A0A7-0E5D538A416A Supplementary Movie 4 D10 cells were seeded in chambered wells in the presence of 8g ml-1 AvSRho (pseudo color) and monitored for 2 hours using a confocal spinning disk microscope to visualize constitutive vesicular exocytosis. Three common cells are shown; data are representative of three impartial experiments. ncomms10823-s5.mov (415K) GUID:?EB2CA1FA-A932-4DF1-BB58-21763A2B87C7 Supplementary Movie 5 The video shows interaction between a peptide pulsed D10 cell expressing CD107a-GFP (pseudo color) and two CTL (previously loaded with Av-SRho, red). Cells 5 were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s6.mov (848K) GUID:?607B7C3F-107E-462E-8F28-210CA3C14D0C Supplementary Movie 6 The video shows interaction between a peptide pulsed D10 cell interacting with a CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s7.mov (689K) GUID:?3720F359-351C-4BEC-BAAE-80AD39FE29F7 Supplementary Movie 7 The video shows interaction between unpulsed D10 cells interacting with CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s8.mov (1.2M) GUID:?8A600A42-61C3-49A5-9ED5-FAFE474E7A4E Supplementary Movie 8 The video shows a peptide pulsed D10 cell interacting with a CTL (that was previously loaded with Av-Alexa488, green) in the presence of 8g ml-1 Av-SRho (red). Cells were inspected using a confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s9.mov (2.8M) GUID:?A276ABD0-8583-48F2-BFD0-9DD109B5C7A8 Supplementary Movie 9 The video shows interactions between a peptide pulsed JY cell and two CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s10.mov (484K) GUID:?FF58E04B-9474-4B05-8C47-404E6CCD5F2D Supplementary Movie 10 The video shows interactions between a peptide pulsed JY cell and a CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s11.mov (394K) GUID:?B4CA3EF7-4205-4980-BD8E-23C6E0FED6D4 Supplementary Movie 11 The video shows interactions between SNAP-23 silenced peptide-pulsed D10 cells expressing CD107a-GFP (pseudo color) and a CTL (previously loaded with AvSRho, red). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner ncomms10823-s12.mov (518K) GUID:?3659F768-DA08-4C83-9127-AF348EE187CA Supplementary Movie 12 The video shows interactions between SNAP-23 silenced peptide-pulsed D10 cells interacting with CTL (previously loaded with CMFDA, green) in the (S)-JQ-35 presence of 8g/ml Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s13.mov (960K) GUID:?8AD91BCA-0A89-4649-839A-456E6B974F83 Supplementary Movie 13 The video shows interactions between control shRNA transfected peptidepulsed D10 cells expressing CD107a-GFP (pseudo color) and a CTL (previously loaded with Av-SRho, red). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s14.mov (800K) GUID:?234FE881-B090-42BA-A434-A2F23A2642F5 Supplementary Movie 14 The video shows interactions between control shRNA transfected peptidepulsed D10 cells interacting with CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s15.mov (579K) GUID:?3511179A-BC9F-4DFB-98D3-2DC711901636 Supplementary Movie 15 The video shows interactions between control shRNA transfected peptidepulsed D10 interacting with CTL (previously loaded with Av-SRho, red) in the presence of 8g ml-1 Av-Alexa488 (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner ncomms10823-s16.mov (479K) GUID:?411D29CA-D652-4215-9158-8DBDCF56FA70 Supplementary Movie 16 The movie shows a 3-D reconstruction of a conjugate formed between a CTL and an untreated melanoma cell. Cells were conjugated for 5 minutes and stained for perforin (green). ncomms10823-s17.mov (836K) GUID:?5DA14835-0AC7-4C5B-B932-EEE4D1B5B69A Supplementary Movie 17 The movie shows a 3-D reconstruction of a conjugate formed between a CTL and a melanoma cell that was pretreated.Jotereau, Nantes, France), EB81-MEL.B, LB3110-MEL and LB2259-MEL.A (isolated from metastatic melanoma patients, kindly provided by Dr P. monitor PI entry. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s3.mov (1.4M) GUID:?A3824B9F-5B28-45E7-B816-DD6BD5319D2F Supplementary Movie 3 JY cells were seeded in chambered wells in the presence of 8g ml-1 Av-SRho (pseudo color) and monitored for 2 hours using a confocal spinning disk microscope to visualize constitutive vesicular exocytosis. A typical cell is shown; data are representative of three impartial experiments. ncomms10823-s4.mov (518K) GUID:?C366B203-5F9E-4150-A0A7-0E5D538A416A Supplementary Movie 4 D10 cells were seeded in chambered wells in the presence of 8g ml-1 AvSRho (pseudo color) and monitored for 2 hours using a confocal spinning disk microscope to visualize constitutive vesicular exocytosis. Three common cells are shown; data are representative of three impartial experiments. ncomms10823-s5.mov (415K) GUID:?EB2CA1FA-A932-4DF1-BB58-21763A2B87C7 Supplementary Movie 5 The video shows interaction between a peptide pulsed D10 (S)-JQ-35 cell expressing CD107a-GFP (pseudo color) and two CTL (previously loaded with Av-SRho, red). Cells 5 were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s6.mov (848K) GUID:?607B7C3F-107E-462E-8F28-210CA3C14D0C Supplementary Movie 6 The video shows interaction between a peptide pulsed D10 cell interacting with a CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s7.mov (689K) GUID:?3720F359-351C-4BEC-BAAE-80AD39FE29F7 Supplementary Movie 7 The video shows interaction between unpulsed D10 cells interacting with CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells were inspected using confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. ncomms10823-s8.mov (1.2M) GUID:?8A600A42-61C3-49A5-9ED5-FAFE474E7A4E Supplementary Movie 8 The IGLC1 video shows a peptide pulsed D10 cell interacting with a CTL (that was previously loaded with Av-Alexa488, green) in the presence of 8g ml-1 Av-SRho (red). Cells were inspected using a confocal spinning disk microscope. Recording time is usually indicated in minutes in upper left corner. (S)-JQ-35 ncomms10823-s9.mov (2.8M) GUID:?A276ABD0-8583-48F2-BFD0-9DD109B5C7A8 Supplementary Movie 9 The video shows interactions between a peptide pulsed JY cell and two CTL (previously loaded with CMFDA, green) in the presence of 8g ml-1 Av-SRho (pseudo color). Cells had been inspected using confocal rotating drive microscope. Recording period can be (S)-JQ-35 indicated in mins in upper remaining part. ncomms10823-s10.mov (484K) GUID:?FF58E04B-9474-4B05-8C47-404E6CCompact disc5F2D Supplementary Film 10 The video displays interactions between a peptide pulsed JY cell and a CTL (previously packed with CMFDA, green) in the current presence of 8g ml-1 Av-SRho (pseudo color). Cells had been inspected using confocal rotating drive microscope. Recording period can be indicated in mins in upper remaining part. ncomms10823-s11.mov (394K) GUID:?B4CA3EF7-4205-4980-BD8E-23C6E0FED6D4 Supplementary Film 11 The video shows interactions between SNAP-23 silenced peptide-pulsed D10 cells expressing CD107a-GFP (pseudo color) and a CTL (previously packed with AvSRho, crimson). Cells had been inspected using confocal rotating drive microscope. Recording period can be indicated in mins in upper remaining part ncomms10823-s12.mov (518K) GUID:?3659F768-DA08-4C83-9127-AF348EE187CA Supplementary Film 12 The video shows interactions between SNAP-23 silenced peptide-pulsed D10 cells getting together with CTL (previously packed with CMFDA, green) in the current presence of 8g/ml Av-SRho (pseudo color). Cells had been inspected using confocal rotating drive microscope. Recording period can be indicated in mins in upper remaining part. ncomms10823-s13.mov (960K) GUID:?8AD91BCA-0A89-4649-839A-456E6B974F83 Supplementary Movie 13 The video displays interactions between control shRNA transfected peptidepulsed D10 cells expressing CD107a-GFP (pseudo (S)-JQ-35 color) and a CTL (previously packed with Av-SRho, reddish colored). Cells had been inspected using confocal rotating drive microscope. Recording period can be indicated in mins in upper remaining part. ncomms10823-s14.mov (800K) GUID:?234FE881-B090-42BA-A434-A2F23A2642F5 Supplementary Movie 14 The video shows interactions between control shRNA transfected peptidepulsed D10 cells getting together with CTL (previously packed with CMFDA, green) in the current presence of 8g ml-1 Av-SRho (pseudo color). Cells had been inspected using confocal rotating drive microscope. Recording period can be indicated in mins in upper remaining part. ncomms10823-s15.mov (579K) GUID:?3511179A-BC9F-4DFB-98D3-2DC711901636 Supplementary Film 15 The video shows.