Figure 7. Bicuculline inhibited L-type... | Rockefeller University Press

Figure 7.

Bicuculline inhibited L-type Ca2+channels to regulate organoid survival.(A) CACNA1C (red) and GFP (LGR5, green) immunofluorescence staining in intestinal crypts. Scale bar, 50 μm. Asterisks indicate CACNA1C +/LGR5+ cell pairs. (B) Examination of the Fluo-4 AM fluorescent signal by confocal microscopy reflecting the variation of intracellular Ca2+ levels in mouse intestinal crypts after 5-FU or irradiation treatment. Scale bar, 100 μm. (C and D) Representative images of C57BL/6 mouse intestinal organoids 2 d after 5-FU (C) or irradiation treatment (D). Organoids were treated with vehicle, muscimol (1 μM), nifedipine (0.1 μM), or nifedipine (0.1 μM) + muscimol (1 μM). Scale bar, 500 μm (upper) or 100 μm (bottom). Arrows indicate dead organoids. In C and D, values are means ± SEM. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 (two-tailed Student’s t test). Data are representative of two (B) or three (A, C, and D) independent experiments.

Bicuculline inhibited L-type Ca ²⁺ channels to regulate organoid survival . (A) CACNA1C (red) and GFP (LGR5, green) immunofluorescence staining in intestinal crypts. Scale bar, 50 μm. Asterisks indicate CACNA1C ⁺/LGR5⁺ cell pairs. (B) Examination of the Fluo-4 AM fluorescent signal by confocal microscopy reflecting the variation of intracellular Ca²⁺ levels in mouse intestinal crypts after 5-FU or irradiation treatment. Scale bar, 100 μm. (C and D) Representative images of C57BL/6 mouse intestinal organoids 2 d after 5-FU (C) or irradiation treatment (D). Organoids were treated with vehicle, muscimol (1 μM), nifedipine (0.1 μM), or nifedipine (0.1 μM) + muscimol (1 μM). Scale bar, 500 μm (upper) or 100 μm (bottom). Arrows indicate dead organoids. In C and D, values are means ± SEM. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 (two-tailed Student’s t test). Data are representative of two (B) or three (A, C, and D) independent experiments.