Vannier for helpful discussion. terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). Raw images of additional experimental replicates for Figure 2, immunofluorescence NS-2028 experiments: This dataset includes additional images from experimental replicates of the images presented in Figure 2. Immunofluorescence staining was performed as described for Figure 2. Images were taken at 40 objective. Bar, 15m. f1000research-7-19822-s0001.tgz (328K) GUID:?96051B39-90AF-4CCB-BD73-491392873974 Copyright : ? 2019 Verraes A et al. Data associated with the article are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). Raw images of immunoprecipitation experiments for Figure 3, immunoprecipitation: Uncropped data from Figure 3 (A) and replicate (C) for VAMP7 NS-2028 immunoprecipitation from Cos-7 cell lysate overexpressing GFP-tagged mouse, rat or human VAMP7 constructs. Uncropped immunoblotting data from Figure 3 (B) and replicate (D) for VAMP7 immunoprecipitation from WT and VAMP7 KO mouse cortex extracts. Antibodies used for immunoprecipitation and subsequent immunoblotting are indicated. Red dashed lines show GFP-VAMP7 protein and cropped region, respectively. IN=Input (50 g in A and C, 100 g in B and D); SN = supernatant after immunoprecipitation; IP = immunoprecipitate; * = GFP-VAMP7; = Absence of band at GFP-VAMP7 size (~50 kDa); ~: immunoglobulins. f1000research-7-19822-s0002.tgz (828K) GUID:?38560019-6101-4360-AD75-7F08F0AD26CD Copyright : ? 2019 Verraes A et al. Data associated with the article are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). Data Availability StatementThe data referenced by this article are under copyright with the following copyright statement: NS-2028 Copyright: ? 2019 Verraes A et al. Data associated with the article are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). http://creativecommons.org/publicdomain/zero/1.0/ Dataset 1. Raw images of experimental replicates for Figure 1, immunoblotting experiments. This dataset includes uncropped blots for all experimental replicates that are represented in Figure 1. Treatments and immunoblot methods were performed as outlined in Figure 1. Blots were probed with indicated anti-VAMP7 antibodies and anti–tubulin antibodies was used as a loading control. (A) Dataset used for Figure 1, with cropped regions in red dashed line. (B) Additional set of raw images of a replicate experiment. Quantification as performed in Figure 1 is shown in lower panel. Note that although signal intensity and background are different within these two replicates, the relative performance of the different tested antibodies remained the same. DOI: http://dx.doi.org/10.5256/f1000research.15707.d221360 28. Dataset 2. Raw images of additional experimental replicates for Figure 2, immunofluorescence experiments. This dataset includes additional images from experimental replicates of the images presented in Figure 2. Immunofluorescence staining was performed as described for Figure 2. Images were taken at 40 objective. Bar, 15m. DOI: http://dx.doi.org/10.5256/f1000research.15707.d234810 29. Dataset 3. Raw images NS-2028 of immunoprecipitation experiments for Figure 3, immunoprecipitation. Uncropped data from Figure 3 (A) and replicate (C) for VAMP7 immunoprecipitation from Cos-7 cell lysate overexpressing GFP-tagged mouse, rat or human VAMP7 constructs. Uncropped immunoblotting data from Figure 3 (B) and replicate (D) for VAMP7 immunoprecipitation from WT and VAMP7 KO mouse cortex extracts. Antibodies used for immunoprecipitation and subsequent immunoblotting are indicated. Red dashed lines show Rabbit Polyclonal to Akt GFP-VAMP7 protein and cropped region, respectively. IN=Input (50 g in A and C, 100 g in B and D); SN = supernatant after immunoprecipitation; IP = immunoprecipitate; * = GFP-VAMP7; = Absence of band at GFP-VAMP7 size (~50 kDa); ~: immunoglobulins. DOI: http://dx.doi.org/10.5256/f1000research.15707.d234809 30. Version Changes Revised.?Amendments from Version 1 We added the stock concentration expressed in mg/ml of each antibody in Table 1. References of the antibodies used were corrected in text and figures. Mislabelling of tables in the text was corrected and knockout replaced invalidation. Comments were added in the text regarding the absence of commercially available antibodies in the immunoprecipitation assays. A paragraph justifying the choice of standard methods instead of antibody-specific optimal protocol has been added to the discussion, although we agree that this choice might favor some antibodies. Figure 2 and Dataset 2 now include a more reliable visual scoring table instead of intensity profile quantification as we agree it might fail to discriminate a typical distribution of VAMP7 NS-2028 (ie: Golgi-like and peripheral vesicles) from an incorrect one (ie: perinuclear-enriched and peripheral diffuse signal, such as.