Fig. 3: Number of HOXA13 + cells rather than cellular expression levels are associated with metaplasia.

Exposure to bile (at pH 7) (a) or to acid (b), in two in vitro model systems of gastroesophageal reflux disease (GERD), marginally induces the expression of HOXA13 from low baseline expression levels in two primary immortalized squamous esophageal cell lines. Error bars represent the 95% CI of the mean. n = 4 independent experiments. For Het1a, two-tailed t-test was used (p = 0.0096 in a, p = 0.0322 in b), for EPC2-hTERT Dunn’s multiple comparisons test in a (control vs. 200 µM, p > 0.99; control vs. 400 µM, p = 0.0112; 200 µM vs. 400 µM, p = 0.14), one-tailed t-test in b, p = 0.0486. c The number of HOXA13-postive cells are increased in Barrett’s esophagus (BE) and intestinal metaplasia (IM) as compared to normal esophagus or stomach tissue. Healthy and BE esophageal samples are derived from the same patients. Mean ± SEM are shown. n = 4 individuals for esophagus, n = 3 for NAG, CAG, n = 4 for IM, n = 1 for GC, n = 6 for healthy colon, n = 11 for CRC. Graphs are based on the analysis of single cell RNA data seq²¹, GSE134520⁸², GSE81861⁸³. Two-tailed t-test was used for esophagus (p = 0.0443) and colon (p = 0.6808), one-way analysis of variance for stomach with Dunnett’s Multiple Comparison Test (p < 0.0001). d HOXA13 expression level per cell is unchanged. Expression level presented in HOXA13⁺ cells only. H healthy, BE Barret’s esophagus, NAG non-atrophic gastritis, CAG chronic atrophic gastritis, IM intestinal metaplasia of stomach, GC early gastric cancer, and CRC colorectal cancer. n = 37 for H, n = 132 for BE, NA for NAG, n = 5 for CAG, n = 163 for IM, n = 69 for GC, n = 37 for healthy colon, n = 87 for CRC of single cells from the individuals mentioned in (c). For GC, statistics are not presented as data per patient was not provided. Boxplots with middle line is the median, the lower and upper hinges correspond to 25th to 75th percentiles, and whiskers representing min-max values.