DOI: 10.5061/dryad.h70rxwdvs
title: Immunostaining images from CRC patients stained with AGR2 and SGTA antibodies

description: The Endoplasmic Reticulum (ER) is an essential
 sensing organelle responsible for the folding and secretion of almost
 one-third of eukaryotic cells' total proteins. However,
 environmental, chemical, and genetic insults often lead to protein
 misfolding in the ER, accumulating misfolded proteins, and causing ER
 stress. To solve this, several mechanisms were reported to relieve ER
 stress by decreasing the ER protein load. Recently, we reported a novel ER
 surveillance mechanism by which proteins from the secretory pathway
 are refluxed to the cytosol to relieve the ER of its content.
  The refluxed proteins gain
 new prosurvival functions in cancer cells, thereby
 increasing cancer cell fitness. We termed this phenomenon ER to CYtosol
 Signaling (or "ERCYS"). Here, we found that in mammalian cells,
 ERCYS is regulated by DNAJB12, DNAJB14, and the HSC70 cochaperone SGTA.
 Mechanistically, DNAJB12 and DNAJB14 bind HSC70 and SGTA - through their
 cytosolically localized J-domains to facilitate ER-protein reflux. DNAJB12
 is necessary and sufficient to drive this phenomenon to increase AGR2
 reflux and inhibit wt-p53 during ER stress. Mutations in DNAJB12/14
 J-domain prevent the inhibitory interaction between AGR2-wt-p53. Thus,
 targeting the DNAJB12/14-HSC70/SGTA axis is a promising strategy to
 inhibit ERCYS  and impair cancer cell fitness.
description: # Immunostaining images from CRC patients stained with AGR2 and SGTA
 antibodies
 [https://doi.org/10.5061/dryad.h70rxwdvs](https://doi.org/10.5061/dryad.h70rxwdvs) ## Description of the data and file structure This dataset is presented in Figure S5F-G as analysed images, here we provide the raw images as well in triplicates. The aim of these images is to provide an evidence that the cytosolic HSC70 co-chaperone colocalizes with  othe ER proteins AGR2 in tumor isolated from Colorectal cancer patients. Those human samples used for the analyses shown in here were provided by the the Colorectal cancer (CRC) biobank in Soroka, Beer Sheva. the samples were collected after Helsinki approval of biobank samples collection, approval number 0137-18. The samples were fixed with 4% paraformaldehyde in PBS for 20 minutes at room temprature. After three washes, samples were permeabilized with  0.5% triton x-100 for 15 min and then washed twice with PBS. 10% goat serum was added for 1hour at room temprature before the addition of the primary antibdies (1) SGTA antibodies (Ptg/60305-1-Ig) and (2) AGR2 antibodies (Ptg/12275-1-AP) overnight at 4°C. samples were washed three times before incubating 1hour with secondary antibodies at room temprature in the dark. After three washes, samples were incubated with 1 μg/ml DAPI and Mounted with a drop of mounting solution ### Files #### File: Control\_1b.czi **Description:**  Images showing the colocalization of SGTA and AGR2 in Healthy tissue (1b) obtained from CRC patients. Each image series, showing SGTA and AGR2 interaction in different sections of the healthy tissue.  #### File: Control\_3.czi **Description:** Images showing the colocalization of SGTA and AGR2 in Healthy tissue (3) obtained from CRC patients. Each image series, showing SGTA and AGR2 interaction in different sections of the healthy tissue.  #### File: Control\_2.czi **Description:** Images showing the colocalization of SGTA and AGR2 in Healthy tissue (2) obtained from CRC patients. Each image series, showing SGTA and AGR2 interaction in different sections of the healthy tissue.  #### File: Tumor1.czi **Description:** Images showing the colocalization of SGTA and AGR2 in Healthy tissue (1) obtained from CRC patients. Each image series, showing SGTA and AGR2 interaction in different sections of the tumor tissue.  #### File: Tumor2.czi **Description:** Images showing the colocalization of SGTA and AGR2 in Healthy tissue (2) obtained from CRC patients. Each image series, showing SGTA and AGR2 interaction in different sections of the tumor tissue.  #### File: Tumor1b.czi **Description:** Images showing the colocalization of SGTA and AGR2 in Healthy tissue (1b) obtained from CRC patients. Each image series, showing SGTA and AGR2 interaction in different sections of the tumor tissue.