Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Feb 4;13(7):905–915. doi: 10.1093/ecco-jcc/jjz010

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2019 European Crohn’s and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com

This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

PMC Copyright notice

Figure 4. — The transcription factor PRDM1 is selectively expressed by CD4⁺ T_RM cells compared with colonic or circulating T_EM cells. A] PRDM1 is selectively expressed by CD4⁺ T_RM cells. CD4⁺ T_RM and colonic and circulating T_EM cells were identified in colonic tissue specimens or from peripheral blood mononuclear cells of Crohn’s disease [CD] patients by flow cytometry as CD69⁺ or CD69^- fractions within the CD4⁺CD45RO⁺CCR7^- compartment. B] Data from [A] are quantified [left panel] and [right panel] the number of PRDM1⁺ cells in each compartment is enumerated [per 1 x 10⁶ total mononuclear cells]. C] PRDM1 inhibition is associated with reduced inflammatory cytokine production by CD4⁺ T_RM cells. CD4⁺ T_RM cells were MACS purified, stimulated with PMA/ionomycin, and co-cultured with the PRDM1 inhibitors butyrate and bortezomib. RNA was extracted and quantitative reverse transcription polymerase chain reaction [qRT-PCR] was performed for the indicated genes. Values were normalised to GAPDH and presented as fold induction relative to the unstimulated condition [negative control]. D] Bortezomib and butyrate reduce inflammatory cytokine production by CD4⁺ T_RM cells in CD. Supernatants from cell cultures from [C] were analysed for production of the indicated cytokines by enzyme-linked immunosorbent assay. E] Bortezomib and butyrate do not affect cell viability. Cells from [C] were counted with tryphan blue to verify viability after stimulation and co-culture, data are presented as the percentage of live cells in the unstimulated condition. Experiments were performed with three biological replicates; *p <0.05, **p <0.01, ***p <0.001, ****p <0.0001 by the unpaired Student’s t test or [D] analysis of variance.P < 0.0001. theunpaired students t-test or the [D] ANOVA.

Figure 4. — The transcription factor PRDM1 is selectively expressed by CD4⁺ T_RM cells compared with colonic or circulating T_EM cells. A] PRDM1 is selectively expressed by CD4⁺ T_RM cells. CD4⁺ T_RM and colonic and circulating T_EM cells were identified in colonic tissue specimens or from peripheral blood mononuclear cells of Crohn’s disease [CD] patients by flow cytometry as CD69⁺ or CD69^- fractions within the CD4⁺CD45RO⁺CCR7^- compartment. B] Data from [A] are quantified [left panel] and [right panel] the number of PRDM1⁺ cells in each compartment is enumerated [per 1 x 10⁶ total mononuclear cells]. C] PRDM1 inhibition is associated with reduced inflammatory cytokine production by CD4⁺ T_RM cells. CD4⁺ T_RM cells were MACS purified, stimulated with PMA/ionomycin, and co-cultured with the PRDM1 inhibitors butyrate and bortezomib. RNA was extracted and quantitative reverse transcription polymerase chain reaction [qRT-PCR] was performed for the indicated genes. Values were normalised to GAPDH and presented as fold induction relative to the unstimulated condition [negative control]. D] Bortezomib and butyrate reduce inflammatory cytokine production by CD4⁺ T_RM cells in CD. Supernatants from cell cultures from [C] were analysed for production of the indicated cytokines by enzyme-linked immunosorbent assay. E] Bortezomib and butyrate do not affect cell viability. Cells from [C] were counted with tryphan blue to verify viability after stimulation and co-culture, data are presented as the percentage of live cells in the unstimulated condition. Experiments were performed with three biological replicates; *p <0.05, **p <0.01, ***p <0.001, ****p <0.0001 by the unpaired Student’s t test or [D] analysis of variance.P < 0.0001. theunpaired students t-test or the [D] ANOVA.