Supplementary Figures S1-3, Tables S1-3 from HIC1 Tumor Suppressor Loss Potentiates TLR2/NF-κB Signaling and Promotes Tissue Damage–Associated Tumorigenesis
Authors: Janeckova Lucie , Pospichalova Vendula , Fafilek Bohumil , Vojtechova Martina , Tureckova Jolana , Dobes Jan , Dubuissez Marion , Leprince Dominique , Baloghova Nikol , Horazna Monika , Hlavata Adela , Stancikova Jitka , Sloncova Eva , Galuskova Katerina , Strnad Hynek , Korinek Vladimir 
Published: April 2023

<p>Supplementary Figures S1-3, Tables S1-3. Supplementary Figure S1. Heatmaps depicting gene expression in Hic1flox/flox MEFs treated with 4- OHT when compared to MEFs treated with vehicle (ethanol) only. Genes (299 in total) displaying significantly (q < 0.05) changed expression in at least one of the indicated time points are listed. The position of Hic1 and six genes whose expression was changed more than twice (|log FC| > 1) in at least two time points is highlighted. Supplementary Figure S2. Loss of Hic1 results in increased counts of goblet and enteroendocrine cells. A,B, goblet (A) and enteroendocrine (B) cell distribution in the indicated segments of the small intestine. Specimens obtained from four Hic1flox/flox and four Hic1flox/flox Villin-Cre+ mice were stained using Periodic acid Schiff (PAS) and an anti-chromogranin A antibody to visualize goblet and enteroendocrine cells, respectively. Stained cells (indicated by black arrowheads in the histology images on the right) were in several different fields indicated by numbers on the X axis. C, no changes in enterocytes were noted in the Hic1-deficient small intestine. Left, qRT-PCR analysis of enterocytespecific markers hairy and enhancer of split-1 (Hes1) and sucrase-isomaltase (SI). The expression level of the respective gene in wt mice (upon normalization to Ubb) was arbitrarily set to 1. Right, staining of brush border enzyme alkaline phosphatase (AP) produced by differentiated enterocytes in the small intestine. Scale bar: 0.15 mm; error bars: SDs. Supplementary Figure S3. DSS-induced transcriptional response in the colon of Hic1flox/flox and Hic1flox/flox Villin-Cre+ mice six and nine days after DSS withdrawal. Epithelial lining of the colon obtained from four animals of each genotype was analyzed using qRT-PCR. The results were normalized to the Ubiquitin B (Ubb) housekeeping gene; the relative expression of another housekeeping gene, β-actin, is also shown. The expression level of the corresponding gene in mice without DSS treatment was arbitrarily set to 1. Error bars: SDs. Supplementary Table S1 Primers used for qRT-PCR analysis Supplementary Table S2. List of genes differentially expressed in Hic1flox/flox MEFs treated with 4- OHT for 48, 72, and 120 hours when compared to control MEFs treated (for the given time periods) with vehicle. Selection criterion: q < 0.05. Of note, none of the genes passed the selection criterion for the 24-hour time point. Supplementary Table S3. The most different 'Gene Ontology Biological Processes' (GO BPs) and WikiPathways categories in expression profiles of Hic1flox/flox MEFs 72 hours after addition of 4-OHT when compared to MEFs treated with vehicle only. In total, 268 gene probes passed the significance criterion (q-value < 0.05). Corresponding annotated genes (listed in Supplementary Table S2) were analyzed using the GO BP and WikiPathways 2015 Enricher datasets. The results were sorted according to the p-value; GO BPs and WikiPathways with p < 0.05 are listed.</p>