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. 2022 Feb 24;13:1030. doi: 10.1038/s41467-022-28569-1

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Heatmap showing all DREAM target genes arranged into ten clusters based on their expression pattern between comparisons among the DEG in hCEcto and hCEcto E6E7 2D stem cells with or without Ctr infection for 48 h. Red frames highlight the cluster containing mismatch repair (MMR) genes. Color bar depicts Z-scored expression values. b Heatmap showing E2F target genes arranged into ten clusters based on their expression pattern between comparisons among the DEG in hCEcto and hCEcto E6E7 2D cells with or without Ctr infection for 48 h. Red frames highlight the cluster containing MMR genes. Color bar depicts log2 FC. c Heatmap depicting the expression of MMR pathway genes. d, e hCEcto and hCEcto E6E7 2D stem cells were infected with Ctr for 48 h. Shown is the relative mRNA expression of MSH6 (d) and MLH1 (e) analyzed by qRT-PCR. Data represent mean ± SDs from three biological replicates normalized to uninfected controls. Statistical significance was calculated using one-way ANOVA, P-values are indicated. f hCEcto and hCEcto E6E7 2D stem cells were infected with Ctr for 48 h, and cell lysates were subjected to immunoblot analysis for indicated proteins and Chlamydia HSP60 and β-actin as a loading control. Data represent four biological replicates. g, h Shown are the hCEcto (g) and hCEcto E6E7 (h) organoids with or without Ctr infection for 5d and immunolabeled for MSH6, Ctr-MOMP, and Nuclei in blue. The arrow highlights the loss of MSH6 expression in hCEcto E6E7. Representative images of three biological replicates. i–k A reporter plasmid containing a G:G mismatch was used for fluorescence-based multiplex flow-cytometric host cell reactivation assay (FM-HCR) to analyze MMR efficiency by the restoration of reporter protein expression. i Comparison of relative reporter expression [%RE] between uninfected and Ctr infected hCEcto 2D stem cells (i) and hCEcto E6E7 (j) cells at 24hpi showing reduced mismatch repair efficiency after infection. Data represent mean ± SDs from two independent experiments normalized to uninfected controls. Source data are provided as a Source Data file.