Fig. 5
FLASH irradiation preserves barrier integrity and enhances tight junction repair in ARPE-19 cells compared to CONV. (A) Bar graph shows Transepithelial Electrical Resistance (TEER) measurements of ARPE-19 cells 7 days post irradiation of increasing intensities, in FLASH (red columns) or CONV (blue columns) modalities. Controls (gray columns) are age-matched culture plates, non-exposed to irradiation. A minimum of 5 independent wells was used for measurements of TEER in each experimental condition. Data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test. Data are expressed as mean ± S.D. p values are indicated in the graphs. (B) Bar graph of qRT-PCR results of ZO-1 gene expression in ARPE-19 cells 24 h and 72 h post irradiation of increasing intensities, either in FLASH (red columns) or CONV (blue columns) modalities. Controls (black dotted line, set to 1) are age-matched culture plates, non-exposed to irradiation. The experiment was conducted on 3 biological replicates for each condition. Data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test and expressed as mean ± SD. Statistically significant results show comparisons between non exposed control cells and irradiated cells. The graphs also show results of comparisons between ARPE-19 irradiated in CONV and ARPE-19 irradiated in FLASH modalities. p values are indicated in the graphs. (C) Representative images of ARPE-19 cells immunostained with ZO-1 antibodies where the intact profile of the tight junctions can be observed in a control culture plate (left, CTRL), while their disruption is evident following oxidative stress (right).
