Figure EV2. ATP‐induced gliosis is mediated by p38‐MAPK and P2X2‐purinergic signaling.

• A
  IL‐6 release measurement by ELISA of IL‐6 in msEGCs. Cells were treated adenosine (1 and 100 µM) or with ATP (100 µM) for 24 h; n = 14–16, msEGCs.
• B
  Protein release measurement by ELISA of IL‐6 in msEGCs. Cells were treated with P2 antagonist PPADS (5, 30 µM) alone or together with ATP (10 or 100 µM) for 24 h; n = 11–12, msEGCs.
• C
  Protein release measurement by ELISA of IL‐6 in msEGCs. Cells were treated with P2X2 antagonist PSB‐1011 (0.2, 2, 20 µM) or PSB‐0711 (0.2, 2, 20 µM) alone or together with ATP (10 µM) for 24 h; n = 9–13, msEGCs.
• D
  Schematic overview of the isolation of msEGCs from small bowel muscularis externa of GFAP^cre‐Ai14^fl/wt mice: FACS‐sorted tdTomato⁺ msEGCs were either analyzed directly (ME‐tissue) or in cultured msEGCs before tdTomato‐FACS‐sorting and further analysis; n = 3–6.
• E
  Gene expression analysis by qPCR of GFAP and Sox10 in msEGC cultures (n = 10) and mouse ME tissue (n = 10).
• F, G
  Representative images of co‐localization of GFAP (green) and tdTomato⁺ msEGC (red) in the ME and in cultured EGCs. Scale bars 50 µm.
• H–K
  qPCR analysis of P2‐purinergic receptors in msEGCs isolated from ME (H, J; n = 3) or from cultured cells (I, K; n = 6), respectively.
• L, M
  Representative Western blots of P2X2 in msEGCs transfected with siRNA‐control or siRNA‐P2X2 for 72 h together with an optical density measurement, see in M. Actin was used as loading control and normalization (n = 6, msEGCs).

Data information: In (A–C and E), data are represented as fold induction + SEM. In (H–K), data are represented as mean + SEM normalized to GAPDH expression. In (M), data are represented as optical density + SEM normalized to actin expression. Statistics were done by applying unpaired Student's t‐test in (A‐C, M and E) or both by unpaired Student's t‐test and one‐way ANOVA with a subsequent Bonferroni test in (B and C). * indicates significance to control, and ^# indicates significance to the ATP treatment with ^# P < 0.05, **/^## P < 0.01, and ***/^### P < 0.001.

Source data are available online for this figure.