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. 2020 Dec 17;13(1):e12724. doi: 10.15252/emmm.202012724

Figure EV5. ExATP induces gliosis in human enteric glia.

Figure EV5

  1. PCA plot of gene expression from patient specimens at two different time points of the surgery; n = 3 for early and late specimens.
  2. IL‐6 protein measurement in human surgical specimens collected during a pancreaticoduodenectomy at an early and a late time point of surgery. Samples were provided on ice directly from the operation room, and muscularis externa (ME) was separated from the lamina propria mucosae; n = 9 human patients.
  3. Gene expression analyses of gliosis marker in human surgical specimens collected during a pancreaticoduodenectomy at an early and a late time point of surgery. The late specimens' mRNA level show an up‐regulation of gliosis genes.
  4. Immunofluorescence microscopy revealed P2X2 expression (green) in a majority of s100β+ (violet) hEGCs in intact myenteric ganglia of the human colon. White arrows mark double‐positive cells. Scale bar 50 µm.
  5. Immunofluorescence microscopy revealed P2X2 expression (green) in a majority of s100β+ (violet) hEGCs in culture. DAPI counterstained nuclei. Quantification of double‐positive cells showed that 75% of cultured hEGCs express P2X2 (marked with +). Scale bar 50 µm.
  6. Schematic workflow on the collection and processing of surgical specimens collected during a pancreaticoduodenectomy. Samples were provided directly from the operation room in oxygenated Krebs–Henseleit buffer and were mechanically activated ex vivo. Immediately after activation, specimens were incubated for 3 h with or without ambroxol (20 µM). Finally, ME was isolated and further processed for qPCR analysis.

Data information: In (B), data are represented as IL‐6 protein in 100 µg tissue; n = 9 human patients. In (C), data are represented as fold change; n = 13 human patients. In (E), data are represented as the percentage of P2X2+/s100β+ cells + SEM; n = 16, hEGCs. Statistics were done by applying unpaired Student's t‐test in (B, C). * indicates significance to control with *P < 0.05, **P < 0.01, and ***P < 0.001.

Source data are available online for this figure.