DOI: 10.5061/dryad.k98sf7mb8
title: High-content imaging, immunoblot and immunofluorescence data related to: Caprin-1 binding to the critical stress granule protein G3BP1 is influenced by pH

description: G3BP is the central node within stress-induced protein–RNA interaction
 networks known as stress granules (SGs). The SG-associated proteins
 Caprin-1 and USP10 bind mutually exclusively to the NTF2 domain of G3BP1,
 promoting and inhibiting SG formation, respectively. Herein, we present
 the crystal structure of G3BP1-NTF2 in complex with a Caprin-1-derived
 short linear motif (SLiM). Caprin-1 interacts with His-31 and His-62
 within a third NTF2-binding site outside those covered by USP10, as
 confirmed using biochemical and biophysical-binding assays.
 Nano-differential scanning fluorimetry revealed reduced thermal stability
 of G3BP1-NTF2 at acidic pH. This destabilization was counterbalanced
 significantly better by bound USP10 than Caprin-1. The G3BP1/USP10 complex
 immunoprecipated from human U2OS cells was more resistant to acidic buffer
 washes than G3BP1/Caprin-1. Acidification of cellular condensates by
 approximately 0.5 units relative to the cytosol was detected by
 ratiometric fluorescence analysis of pHluorin2 fused to G3BP1. Cells
 expressing a Caprin-1/FGDF chimera with higher G3BP1-binding affinity had
 reduced Caprin-1 levels and slightly reduced condensate sizes. This
 unexpected finding may suggest that binding of the USP10-derived SLiM to
 NTF2 reduces the propensity of G3BP1 to enter condensates.
description: The deposited ZIP folders are described in following
 groups: @HTimages_1 high-throughput
 imaging datasets of cellular GFP-G3BP condensation assays presented in
 Figure 5 of the manuscript ZIP folders:
 img_plate2171.zip / img_plate2185.zip / img_plate21691.zip
 @HTimages_2<strong>
 </strong> high-throughput imaging datasets of
 cellular GFP-Caprin condensation assays presented in Figure 7 of the
 manuscript. ZIP folders: img_plate1734.zip /
 img_plate1735.zip / img_plate1737.zip / img_plate1740.zip
 @HTimages_3 
 high-throughput imaging datasets of in-vitro reconstituted
 GFP-G3BP condensation assays (LLPS), presented in Figure 5 of the
 manuscript. ZIP folders:
 LLPS_GFP-G3BP1.zip 
 @HTimages_4 high-throughput imaging
 datasets of in-vitro reconstituted GFP-Caprin condensation assays (LLPS),
 presented in Figure 7 of the manuscript. ZIP folders:
 LLPS_GFP-Caprin-1.zip 
 @CPpipelines_1 Cellprofiler pipelines to
 analyse @HTimages_1  ZIP folders: pipelines_G3BP1.zip
 t @CPpipelines_2 
 Cellprofiler pipelines to analyse @HTimages_2 
 ZIP folders:pipelines_Caprin1.zip 
 @Images_1 raw image files used
 for presentation of Blots or IF data in main and supplemental
 figures ZIP folders: Images_Blots.zip
 The collection of
 datasets is described in the methods section of the associated manuscript
 (ms, https://royalsocietypublishing.org/doi/10.1098/rsob.220369) @HTimages_1/@HTimages_2: <em>ms section: 4.9.1. High-content microscopy</em>:  "Images were recorded with a Molecular Devices ImageXpress Micro microscope, equipped with a 20x or 40x objective, and illuminated with a mercury lamp and standard filters for DAPI, FITC, Cy3 and Cy5. Images were captured using a four-megapixel sCMOS digital camera with the manufacturer’s software MetaXpress, and raw TIF files were analyzed using CellProfiler (CP), ImageJ and Rstudio." @HTimages_3/@HTimages_4 <em>ms section: </em>4.9.4. In vitro reconstituted condensate assays "Images were taken with a Supercontinuum Confocal Leica TCS SP5 X, equipped with a pulsed white light laser and a Leica HCX PL Apo 63x/1.40 oil objective. "<strong><br></strong> @Images_1 <em>ms section </em>4.8. Immunoprecipitation and immunoblotting <em>ms section: </em>4.9. Immunofluorescence analysis
description: See README file for full details. 
 general notes on deposited file formats:<br>*.TIF or *.tif
 image files can be opened using standard software for images (ImageJ,
 Photoshop,...)<br>*.xlsx can be opened using Microsoft
 Excel<br>*.cpproj project files can be opened in Cellprofiler (https://cellprofiler.org/) High-throughput (HT) imaging datasets can be analysed by opening the deposited Cellprofiler pipelines in Cellprofiler (https://cellprofiler.org/).<br>TIF files can also be viewed using any image viewing software (https://imagej.nih.gov/ij/download.html). <em>Scripts to analyse the Cellprofiler output, and additional information is found on Github:</em> https://github.com/derpaule/RSOB-22-0369  
description: # Supplementary data to rsob.220369: high-throughput imaging datasets and
 western blot images. ## Overview of uploaded ZIP folders In this
 repository following ZIP folders were deposited. Listed with NAME of the
 ZIP-folder, SIZE, and a group-TAG. Analysis scripts related to the folders
 are available on GITHUB | NAME | SIZE | TAG | GITHUB | | ---- | ---- | ---
 | ------ | | Images\_Blots.zip | 216.08 MB | @Images\_1 | - | |
 pipelines\_G3BP1.zip | 430.17 KB | @CPpipelines\_1 |
 https://derpaule.github.io/RSOB-22-0369\_condensates\_GFP-G3BP1 | |
 img\_plate2171.zip | 2.77 GB | @HTimages\_1 |
 https://derpaule.github.io/RSOB-22-0369\_condensates\_GFP-G3BP1 | |
 img\_plate2185.zip | 3.47 GB | @HTimages\_1 |
 https://derpaule.github.io/RSOB-22-0369\_condensates\_GFP-G3BP1 | |
 img\_plate21691.zip | 3.27 GB | @HTimages\_1 |
 https://derpaule.github.io/RSOB-22-0369\_condensates\_GFP-G3BP1 | |
 img\_plate1734.zip | 1.09 GB | @HTimages\_2 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-Caprin1 | |
 img\_plate1735.zip | 1.12 GB | @HTimages\_2 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-Caprin1 | |
 img\_plate1737.zip | 1.16 GB | @HTimages\_2 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-Caprin1 | |
 img\_plate1740.zip | 1.09 GB | @HTimages\_2 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-Caprin1 | |
 pipelines\_Caprin1.zip | 473.67 KB | @CPpipelines\_2 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-Caprin1 | |
 LLPS\_GFP-G3BP1.zip | 320.97 MB | @HTimages\_3 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-G3BP1 | |
 LLPS\_GFP-Caprin-1.zip | 181.39 MB | @HTimages\_4 |
 https://github.com/derpaule/RSOB-22-0369\_condensates\_GFP-Caprin1 | ##
 GENERAL NOTES ### notes on tag groups: `@HTimages_1` group comprises
 high-throughput imaging datasets of cellular GFP-G3BP condensation assays
 presented in Figure 5 of the manuscript. `@HTimages_2` group comprises
 high-throughput imaging datasets of cellular GFP-Caprin condensation
 assays presented in Figure 7 of the manuscript. `@HTimages_3` group
 comprises high-throughput imaging datasets of in-vitro reconstituted
 GFP-G3BP condensation assays (LLPS), presented in Figure 5 of the
 manuscript. `@HTimages_4` group comprises high-throughput imaging datasets
 of in-vitro reconstituted GFP-Caprin condensation assays (LLPS), presented
 in Figure 7 of the manuscript. `@CPpipelines_1` group comprises
 Cellprofiler pipelines to analyse @HTimages\_1 `@CPpipelines_2` group
 comprises Cellprofiler pipelines to analyse @HTimages\_2 `@Images_1` group
 comprises raw image files used for presentation of Blots or IF data in
 main and supplemental figures ### notes on deposited file formats: `.TIF`
 or `*.tif` image files can be opened using standard software for images
 (ImageJ, Photoshop,...) `.xlsx` can be opened using Microsoft Excel
 `.cpproj` project files can be opened in
 [Cellprofiler](https://cellprofiler.org/), `@CPpipelines_1` are associated
 with `@HTimages_1`, `@CPpipelines_2` are associated with `@HTimages_2`;
 `@HTimages_3` are analysed by
 `21-08-16_LLPScondensatecount_GFPimage.cpproj` ### structure of FILELIST
 files are organised as follows: #### ZIPfolder * **folder** * `filename` *
 filename description or placeholder (see below) ### filename patterns in
 FILELIST Filename pattern `[A|B]_[1-10].tif` would result in 20 tif image
 files: * A\_1.tif ... A\_10.tif * B\_1.tif ... B\_10.tif `*` in filename
 patterns was used as regular expression: `*Mock*.tif` would correspond to
 all TIF files containing the word "Mock" The original image
 filenames were overly long and often comprised empty spaces or comma
 signs. As requested by the curators, the filenames were shortened compared
 to the original filenames. However, the uploaded cell profiler scripts or
 Excel sheets may refer to the old filenames. To facilitate identification
 of the renamed files, we refer to the original filename as follows, using
 placeholders: as i.e. `2Y` = `SG 40x ddG3- G1-wt- H31A- 2Y` If the
 shortened filenames comprised spaces ` ` or comma signs `,` these were
 substituted by `x` or `X` and these changes are labeled as `/ /x/` and
 `/,/X/` respectively ## SPECIFIC NOTES TO TAG GROUPS ### @HTimages\_1 and
 @HTimages\_2 comprise high-throughput (HT) imaging datasets that can be
 analysed by opening the associated pipelines *@CPpipelines\_1* and
 *@CPpipelines\_2* in [Cellprofiler](https://cellprofiler.org/) TIF files
 can also be viewed using any image viewing software
 [ImageJ](https://imagej.nih.gov/ij/download.html). #### specifics to
 filename pattern: `[A01|B01|C01]` identifiers for cell-lines expressing
 different G3BP or Caprin-1 variants, specified in deposited pipelines
 *@CPpipelines\_1* and *@CPpipelines\_2* `[s1-s100]` recorded wells for
 each cell-line `w[1-2]/w3` each well was measured at three wavelengths ###
 @HTimages\_3 and @HTimages\_4 datasets comprise high-throughput (HT)
 imaging datasets that can be analysed by opening the associated pipeline
 "21-08-16\_LLPScondensatecountGFPimage.cpproj" in
 [Cellprofiler](https://cellprofiler.org/) TIF files can also be viewed
 using any image viewing software
 (https://imagej.nih.gov/ij/download.html). #### specifics to filename
 pattern: `[2Y|H31A|wt]` identifiers for cell-lines expressing different
 G3BP variants, as specified in manuscript `[wt|FGDF]` identifier for
 Caprin-1 variant `[5|10|15|20|25|30]` measured concentrations in
 micromolar `[1|2|3]` number of replicate `ch0[0|1]` recorded channel;
 channel 0 is the relevant channel for GFP ### @Images\_1 raw images used
 for the figures presented in the manuscript, organised in folder and
 subfolders according to Figures in manuscript. ## FILELIST ###
 @HTimages\_1 #### img\_plate2171.zip * **plate\_01**: *1200* TIF image
 files * `2Y_[A01|B01|B02|B03]_[s1-s100]_w[1-2].TIF` *
 `2Y_[A01|B01|B02|B03]_[s1-s100]_w3.tif` * with placeholder: `2Y` = `SG 40x
 ddG3- G1-wt- H31A- 2Y` * **plate\_02**: *1200* TIF image files with
 following naming pattern: * `2Y_[C01|D01|D02|D03]_[s1-s100]_w[1-2].TIF` *
 `2Y_[C01|D01|D02|D03]_[s1-s100]_w3.tif` * with placeholder: `2Y` = `SG 40x
 ddG3- G1-wt- H31A- 2Y` #### img\_plate2185.zip * **plate\_01**: *1200* TIF
 image files with following * `2Y_[A01|B01|B02|B03]_[s1-s100]_w[1-2].TIF` *
 `2Y_[A01|B01|B02|B03]_[s1-s100]_w3.tif` * with placeholder: `2Y` = `SG 40x
 ddG3- G1-wt- H31A- 2Y` * **plate\_02**: *1200* TIF image files with
 following * `2Y_[C01|D01|D02|D03]_[s1-s100]_w[1-2].TIF` *
 `2Y_[C01|D01|D02|D03]_[s1-s100]_w3.tif` * with placeholder: `2Y` = `SG 40x
 ddG3- G1-wt- H31A- 2Y` #### img\_plate21691.zip * **plate\_01**: *1200*
 TIF image files with following *
 `2Y_[A01|B01|B02|B03]_[s1-s100]_w[1-2].TIF` *
 `2Y_[A01|B01|B02|B03]_[s1-s100]_w3.tif` * with placeholder: `2Y` = `SG 40x
 ddG3- G1-wt- H31A- 2Y` * **plate\_02**: *1200* TIF image files with *
 `2Y_[C01|D01|D02|D03]_[s1-s100]_w[1-2].tif` *
 `2Y_[C01|D01|D02|D03]_[s1-s100]_w3.TIF` * with placeholder: `2Y` = `SG 40x
 ddG3- G1-wt- H31A- 2Y` ### @HTimages\_2 #### img\_plate1734.zip *
 **1734\_A01**: *300* TIF image files with following naming pattern: -
 `FGDF_A01_[s1-s100]_w[1-2].tif` - `FGDF_A01_[s1-s100]_w3.TIF` *
 **1734\_A02**: *300* TIF image files with following naming pattern: -
 `FGDF_A02_[s1-s100]_w[1-2].tif` - `FGDF_A02_[s1-s100]_w3.TIF` *
 **1734\_B01**: *300* TIF image files with following naming pattern: -
 `FGDF_B01_[s1-s100]_w[1-2].tif` - `FGDF_B01_[s1-s100]_w3.TIF` *
 **1734\_B02**: *300* TIF image files with following naming pattern: -
 `FGDF_B02_[s1-s100]_w[1-2].tif` - `FGDF_B02_[s1-s100]_w3.TIF` * with
 placeholder: `FGDF` = `SG 40x Cap1-wt FGDF` #### img\_plate1735.zip *
 **1735\_A01**: *300* TIF image files with following naming pattern: *
 `FGDF_A01_[s1-s100]_w[1-2].tif` * `FGDF_A01_[s1-s100]_w3.TIF` *
 **1735\_A02**: *300* TIF image files with following naming pattern: *
 `FGDF_A02_[s1-s100]_w[1-2].tif` * `FGDF_A02_[s1-s100]_w3.TIF` *
 **1735\_B01**: *300* TIF image files with following naming pattern: *
 `FGDF_B01_[s1-s100]_w[1-2].tif` * `FGDF_B01_[s1-s100]_w3.TIF` *
 **1735\_B02**: *300* TIF image files with following naming pattern: *
 `FGDF_B02_[s1-s100]_w[1-2].tif` * `FGDF_B02_[s1-s100]_w3.TIF` * with
 placeholder: `FGDF` = `SG 40x Cap1-wt FGDF` #### img\_plate1737.zip *
 **1737\_A01**: *300* TIF image files with following naming pattern: *
 `FGDF_A01_[s1-s100]_w[1-2].tif` * `FGDF_A01_[s1-s100]_w3.TIF` *
 **1737\_A02**: *300* TIF image files with following naming pattern: *
 `FGDF_A02_[s1-s100]_w[1-2].tif` * `FGDF_A02_[s1-s100]_w3.TIF` *
 **1737\_B01**: *300* TIF image files with following naming pattern: *
 `FGDF_B01_[s1-s100]_w[1-2].tif` * `FGDF_B01_[s1-s100]_w3.TIF` *
 **1737\_B02**: *300* TIF image files with following naming pattern: *
 `FGDF_B02_[s1-s100]_w[1-2].tif` * `FGDF_B02_[s1-s100]_w3.TIF` * with
 placeholder: `FGDF` = `SG 40x Cap1-wt FGDF` #### img\_plate1740.zip *
 **1740\_A01**: *300* TIF image files with following naming pattern: *
 `FGDF_A01_[s1-s100]_w[1-2].tif` * `FGDF_A01_[s1-s100]_w3.TIF` *
 **1740\_A02**: *300* TIF image files with following naming pattern: *
 `FGDF_A02_[s1-s100]_w[1-2].tif` * `FGDF_A02_[s1-s100]_w3.TIF` *
 **1740\_B01**: *300* TIF image files with following naming pattern: *
 `FGDF_B01_[s1-s100]_w[1-2].tif` * `FGDF_B01_[s1-s100]_w3.TIF` *
 **1740\_B02**: *300* TIF image files with following naming pattern: *
 `FGDF_B02_[s1-s100]_w[1-2].tif` * `FGDF_B02_[s1-s100]_w3.TIF` * with
 placeholder: `FGDF` = `SG 40x Cap1-wt FGDF` ### @CPpipelines\_1 ####
 pipelines\_G3BP1.zip * *6* [Cellprofiler](https://cellprofiler.org/)
 pipelines to analyse @Images\_1 *
 `SG_pipeline04_caprin_plate2169_01_GFPint02.cpproj` *
 `SG_pipeline04_caprin_plate2169_02_GFPint02.cpproj` *
 `SG_pipeline04_caprin_plate2171_01_GFPint02.cpproj` *
 `SG_pipeline04_caprin_plate2171_02_GFPint02.cpproj` *
 `SG_pipeline04_caprin_plate2185_01_GFPint02.cpproj` *
 `SG_pipeline04_caprin_plate2185_02_GFPint02.cpproj` ### @CPpipelines\_2
 #### pipelines\_Caprin1.zip * *5*
 [Cellprofiler](https://cellprofiler.org/) pipelines to analyse @Images\_2
 * `SG_pipeline_chimera_1734_robust.cpproj` *
 `SG_pipeline_chimera_1735_robust.cpproj` *
 `SG_pipeline_chimera_1737_robust.cpproj` *
 `SG_pipeline_chimera_1740_robust_CytoNorm.cpproj` *
 `SG_pipeline_chimera_1740_robust.cpproj` ### @HTimages\_3 ####
 LLPS\_GFP-G3BP1.zip * `21-08-16_LLPScondensatecount_GFPimage.cpproj`
 Cellprofiler pipeline to analyse the images of @HTimages\_3 *
 **21-11-02\_LLPS\_2Y** *
 `LLPS_2Y_[2Y|H31A|wt]XxrG1-[2Y|H31A|wt]Xx[5|10|15|20|25|30]xuM-[1|2|3]_z0_ch0[0|1].tif` * *108* TIF images of G3BP\-\[WT\|2Y\|H31A\] spiked in at six concentrations\, measured in triplicate\, in two channels * with placeholders: `LLPS_2Y_` = `21-11-02, LLPS, GFP-G1wt, H31A, 2Y, rG1-wt, H31A, 2Y, 30-5 uM, 1h_GFP-G1-` + `/ /x/` + `/,/X/` * `21-11-02_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-11-03\_LLPS\_2Y** * `LLPS_2Y_[2Y|H31A|wt]XxrG1-[2Y|H31A|wt]Xx[0|5|10|15|20|25|30]xuM-[1|2|3]_z0_ch0[0|1].tif` * *126* TIF images of G3BP\-\[WT\|2Y\|H31A\] spiked in at six concentrations\, measured in triplicate\, in two channels * with placeholder: `LLPS_2Y_` = `21-11-03, LLPS, GFP-G1wt, H31A, 2Y, rG1-wt, H31A, 2Y, 30-5 uM, 1h_GFP-G1-` + `/ /x/` + `/,/X/` * `21-11-04_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-12-03\_LLPS\_2Y** * `LLPS_2Y_[2Y|H31A|wt]XxrG1-[2Y|H31A|wt]Xx[0|5|10|15|20|25|30]xuMXx1h-[1|2|3]_z0_ch0[0|1].tif` * *126* TIF images of G3BP\-\[WT\|2Y\|H31A\] spiked in at six concentrations\, measured in triplicate\, in two channels * with placeholder: `LLPS_2Y_` = `21-12-03, LLPS, GFP-G1wt, H31A, 2Y, rG1-wt, H31A, 2Y, 30-0 uM, 1h_GFP-G1-` + `/ /x/` + `/,/X/` * `21-12-03_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-12-10\_LLPS\_2Y** * `LLPS_2Y_[2Y|H31A|wt]XxrG1-[2Y|H31A|wt]Xx[0|5|10|15|20|25|30]xuM-[1|2|3]_z0_ch0[0|1].tif` * *126* TIF images of G3BP\-\[WT\|2Y\|H31A\] spiked in at six concentrations\, measured in triplicate\, in two channels * with placeholder: `LLPS_2Y_` = `21-12-10, LLPS, GFP-G1wt, H31A, 2Y, rG1-wt, H31A, 2Y, 30-0 uM_GFP-G1-` + `/ /x/` + `/,/X/` * `21-12-10_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet ### @HTimages\_4 #### LLPS\_GFP-Caprin-1.zip * **21-08-17\_LLPS\_FGDF** * `LLPS_FGDF_[wt|FGDF]XxrG1-wtx[1.25|2.5|5|10|20]xuMXx1h-[1|2|3]_z0_ch0[0|1].tif` * *60* TIF images of GFP\-Cap1\-\[wt\|FGDF\] spiked in at 5 concentrations in triplicate measured in two channels * with placeholders: `LLPS_FGDF_` = `21-08-17, LLPS, GFP-Cap1-wt vs FGDF 5 mg-mL, rG1-wt_LLPS, GFP-Cap1-` + `/ /x/` + `/,/X/` * `21-08-17_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-08-23\_LLPS\_FGDF** * `LLPS_FGDF_[wt|FGDF]XxrG1-wtx[1.25|2.5|5|10|20]xuMXx1h-[1|2|3]_z0_ch0[0|1].tif` * *60* TIF images of GFP\-Cap1\-\[wt\|FGDF\] spiked in at 5 concentrations in triplicate measured in two channels * with placeholders: `LLPS_FGDF_` = `21-08-23, LLPS, GFP-Cap1-wt vs FGDF, 5 mg-ml, rG1-wt_LLPS, GFP-Cap1-` + `/ /x/` + `/,/X/` * `21-08-23_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-08-27\_LLPS\_FGDF** * `LLPS_FGDF_[wt|FGDF]XxrG1-wtx[1.25|2.5|5|10|20]xuMXx1h-[1|2|3]_z0_ch0[0|1].tif` * *60* TIF images of GFP\-Cap1\-\[wt\|FGDF\] spiked in at 5 concentrations in triplicate measured in two channels * with placeholders: `LLPS_FGDF_` = `21-08-27, LLPS, GFP-Cap1-wt vsFGDF, 5 mg-mL, rG1-wt 1h_LLPS, GFP-Cap1-` + `/ /x/` + `/,/X/` * `21-08-27_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-12-13\_LLPS\_FGDF** * `LLPS_FGDF_[wt|FGDF]XxrG1-wtXx[0|5|10|15|20|25|30]xuM-[1|2|3]_z0_ch0[0|1].tif` * *84* TIF images of GFP\-Cap1\-\[wt\|FGDF\] spiked in at 7 concentrations in triplicate measured in two channels * with placeholders: `LLPS_FGDF_` = `21-12-13, LLPS, GFP-Cap1-wt vs FGDF, 4.2 mg, rG1-wt 30-0 uM_GFP-Caprin1-`+ `/ /x/` + `/,/X/` * `21-12-13_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet * **21-12-23\_LLPS\_FGDF** * `LLPS_FGDF_[wt|FGDF]XxrG1x[0|5|10|15|20|25|30]xuM-2_z0_ch0[0|1].tif` * *84* TIF images of GFP\-Cap1\-\[wt\|FGDF\] spiked in at 7 concentrations in triplicate measured in two channels * with placeholder + substitutions: `LLPS_FGDF_` = `21-12-23, LLPS, GFP-Cap wt vs FGDF, rG1 0-30 uM, 1h_GFP-Cap1-` + `/ /x/` + `/,/X/` * `21-12-23_Analysis_Coverage.xlsx` Analysis of Cellprofiler output in Excel sheet ### @Images\_1 #### Images\_Blots.zip * **MainFigures** * **Figure2** * **Figure2A\_westernblots** * `2s_IPs360-370_Cap1_Actin_G1_G2.tif` * `10s_Cap1_GAPDH_G1rerunlysates.tif` * `10s_Cap1_GAPDH_G1_lysates.tif` * `10s_IPs371-381_Cap1_G1_G2.tif` * `30s_Cap1_GAPDH_G1_lysates.tif` * `180s_IPs360-370_Cap1_Actin_G1_G2.tif` * `300s_IPs371-381_Cap1_G1_G2.tif` * `500s_IPs371-381_Cap1_G1_G2.tif` * `white_Cap1_GAPDH_G1rerunlysates.tif` * `white_Cap1_GAPDH_G1_lysates.tif` * `white_IPs360-370_Cap1_Actin_G1_G2.tif` * `white_IPs371-381_Cap1_G1_G2.tif` * **Figure4** * **Figure4B\_westernblots** * `WB_1s_G1_Cap1_USp10_GAPDH.tif` * `WB_5s_UBAP2L_USP10_Cap1_G1_GAPDH_RPS6.tif` * `WB_10s_G1_Cap1_USp10_GAPDH.tif` * `WB_20s_UBAP2L_USP10_Cap1_G1_GAPDH_RPS6.tif` * `WB_60s_UBAP2L_USP10_Cap1_G1_GAPDH_RPS6.tif` * `WB_480s_UBAP2L_USP10_Cap1_G1_GAPDH_RPS6.tif` * `WB_white_G1_Cap1_USp10_GAPDH.tif` * `WB_white_UBAP2L_USP10_Cap1_G1_GAPDH_RPS6.tif` * **Figure5** * **Figure5A\_IFimages** * `2Y_A01_s56_w[1|2|3].tif` *3* TIF images * `2Y_B01_s100_w[1|2|3].tif` *3* TIF images * `2Y_B02_s20_w[1|2|3].tif` *3* TIF images * `2Y_B03_s53_w1[1|2|3].tif` *3* TIF images * \*\*Figure5D\_IFimages * `LLPS_2Y_[2Y|H31A|wt]XxrG1-[2Y|H31A|wt]Xx[0|5|10|15|20|25|30]xuM-1_z0_ch0[0|1].tif` *42* TIF images of G3BP\-\[WT\|2Y\|H31A\] spiked in at six concentrations\, two channels * **Figure6** * **Figure6A\_IFimages** * `ddG32_z0_ch00.tif` * `ddG32_z0_ch01.tif` * \*\*Figure6D\_IFimages * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c1_green.tif` * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c1-3.tif` * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c1-c3.tif` * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c1.tif` * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c2.tif` * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c3_blue.tif` * `22-01-112nd_Phlu2-G1rG1pH7_4-4-ImageExport-10_c3.tif` * **Figure7** * **Figure7A\_westernblot** * `WB_10s_G1_GAP_FMR_Cap1.tif` * `wb_500s_O.N.FMR1.tif` * `WB_white_G1_GAP_FMR_Cap1.tif` * `wb_white_O.N.FMR1.tif` * **Figure7C\_IFimages** * `FGDF_A01_s10_w[1|2|3].tif` *3* TIF images * `FGDF_A02_s9_w[1|2|3].tif` *3* TIF images * `FGDF_B02_s9_w[1|2|3].tif` *3* TIF images * **Figure7F\_IFimages** * `LLPS_FGDF_[wt|FGDF]XxrG1-wtXx[0|5|10|15|20|25|30]xuM-1_z0_ch0[0|1].tif` *28* TIF images of GFP\-Cap1\-\[wt\|FGDF\] spiked in at 7 concentrations in triplicate measured in two channels * \*\* SupplementalFigures \*\* * **SupplementalFigureS3** * **SupplementalFigureS3B\_westernblot** * `15-05-12_PoncU2OSCaprin1KO.tif` * `240s_UBAP2L_G1_Cap1_GAPDH.tif` * `white_UBAP2L_G1_Cap1_GAPDH.tif` * **SupplementalFigureS3D\_IFimages** * `U2OS-wt+U2OS-Cap1-KOg2_MockCap1-2.tif` * `U2OS-wt+U2OS-Cap1-KOg2_MockG3BP1.tif` * `U2OS-wt+U2OS-Cap1-KOg2_MockTIA1.tif` * `U2OS-wt+U2OS-Cap1-KOg2_SACap1-2.tif` * `U2OS-wt+U2OS-Cap1-KOg2_SAG3BP1.tif` * `U2OS-wt+U2OS-Cap1-KOg2_SATIA1.tif` * **SupplementalFigureS6** * `10s_IPs_UBAP2L_G1_Cap1_USP10_GAPDH.tif` * `10s_Lys_G3BP1reblot(good).tif` * `30s_IPs_UBAP2L_G1_Cap1_USP10_GAPDH.tif` * `240s_Lys_UBAP2L_G1_Cap1_GAPDH.tif` * `white_IPs_UBAP2L_G1_Cap1_USP10_GAPDH.tif` * `white_Lys_UBAP2L_G1_Cap1_GAPDH.tif` * **SupplementalFigureS8** * `H31A_C02_s10_w1-merge-rgb.tif` * `H31A_C02_s10_w[1|2|3]-rgb.tif` *3* TIFs * `ddG_G1-wt-H31A-2Y_B01_s10_w3merge-rgb.tif` * `ddG_G1-wt-H31A-2Y_B01_s10_w[1|2|3]-rgb.tif` *3* TIFs * **SupplementalFigureS9** * `LLPS_FGDF_[2y|H31A|wt]XxrG1-[2Y|H31A|wt]Xx20xuMXx[6.8|7.0|7.2|7.4|7.6|7.8]-1_z0_ch0[0|1].tif` *36* TIF images