DOI: 10.5281/zenodo.16676359
title: Localization of PRC1 subunits relative to Hox chromatin in super-resolution microscopy during Drosophila embryogenesis

description: The images contained in this database are associated to the paper: “PRC1 nano-structures compact Hox chromatin during Drosophila embryogenesis.”

 

A full description of the methods used to perform the experiments and how the images were acquired is described in the “method” section of this article. Briefly, this work was entirely carried out on Drosophila embryos, collecting images from the outermost cell layer. The embryonic stages and the location where images were acquired are mentioned in the name of each file. The Excel file “PRC1_NG_table1” compiles all the raw data files (highlight in yellow) used in this work and clearly indicates their stage and location.

In this work, we had detected 3 subunits of PRC1 through immuno-labellings:

·       Ph for Polyhomeotic

·       Pc for Polycomb

·       Psc for Posterior Sex Comb

 

To compare the nuclear localization of these proteins, we performed immuno-FISH experiments with probes recognizing these sequences of DNA (Dm6 coordinates):

·       eUbx: 16616022-16621977

·       bx: 16700036-16705661

·       int1: 16737398-16743027

·       bxd: 16762746-16768281

·       int2: 16785088-16790830

·       Fab7: 16896724-16902352

·       eAbdB: 16983104-16988951

·       dAntp: 7032521-7038481

·       int3: 7016161-7021673

·       Antp: 6998210-7003802

·       Int4: 6949915-6955302

·       Scr: 6846535-6852075

·       elab: 6632112-6637915

·       Cont1: 17301755-17307166

·       Cont2: 14977118-14982758

 And full BX-C: 16660000-16975000

 

This database is composed of 8 folders compressed with 7-Zip and containing the raw data of the images used in this work:

·       Immuno_FISH_Ph, Immuno_FISH_Pc and Immuno_FISH_PcXT109 contain images in .msr format which is the Abberior format directly produced by the Abberrior STED microscope. Ph or Pc immuno-labelling used the STAR-RED fluorochrome and FISH probes used the Alexa 594 fluorochrome. Pixel size is 20 nm.

·       Immuno_Ph and Immuno_Pc contain images in .tif format with channel 1 corresponding to confocal images and channel 2 corresponding to STED images. Ph or Pc immunolabellings are detected with the STAR-RED fluorochrome and images have a pixel size of 20 nm. Stacks consist of 5 optical sections spaced by 300 nm.

·       Immuno_PcXT109 contain images in .tif format with channel 1 corresponding to confocal images of Ph, channel 2 corresponding to STED images of Ph and channel 3 corresponding to confocal images of Pc. Ph is detected with the STAR-RED fluorochrome whereas Pc used the Alexa 594 fluorochrome. Pixel size is 20 nm.

·       Coloc_Pc_Ph_FigS1C contain images in .tif format with channel 1 (C1) corresponding to Ph in STED, channel 2 (C2) corresponding to Pc in STED, channel 3 (C3) corresponding to Ph in confocal and channel 4 (C4) corresponding to Pc in confocal. This file contains all the data used to quantify Ph and Pc co-localization and correspond to results presented in Fig. S1C.

·       Image_ms contain images in .tif format, except original Airy Scan images (Fig. 1E and Fig. S4) which are in .czi format (Airy Scan microscope is a Zeiss microscope producing  .czi files). This file contains all the images shown in the manuscript and their associated raw data.