Fig. 6

Radiation induce phosphorylation of histone H2A.X. a Confocal microscopic images of rectal organoids exposed to 0, 2, 4, 6, and 8 Gy of fractionated radiation. Increase in histone phosphorylation (γH2A.X green color) was observed in a dose-dependent manner. b Histogram demonstrating significant increase in relative fluorescence units (RFU) representing immune-fluorescence signal in organoids exposed to 8 Gy of fractionated radiation, (n = 4, p value < 0.05, Mann-Whitney test) compared to un-irradiated control. c Confocal microscopic images of rectal organoids treated with doxorubicin (1 μM). Note the increase in γH2A.X (green color) in doxorubicin-treated organoids compared to control. d Histogram demonstrating significant increase in relative fluorescence units (RFU) representing immune-fluorescence signal in organoids exposed to doxorubicin (n = 3, p value < 0.001, Mann-Whitney test) compared to un-irradiated control. e, f Microscopic image (phase contrast) of rectal organoids along with quantification of % of budding organoid demonstrating that doxorubicin impaired the organoid growth compared to un-irradiated control (p < 8.9217E−05). g Histogram demonstrating significant decrease in organoid viability with doxorubicin treatment compared to control (p < 1.57794E−05)