Prolonged exposure to intracellular environment analysis was performed after washing at time t. Here, QSH or PS washed samples at time t were left to incubate to an additional 12 − x and 24 − x hours, where x is the time of wash for each particular sample. At total experimental time of 12 and 24 hours, previously washed plates were mixed and measured for fluorescence changes from their original time t. An example is shown below:

2-hour wash fluorescence ➔ 10-hour post-wash (12 2 hours) fluorescence ➔ 22-hour post-wash (24 2 hours) fluorescence.

Twelve-hour washed sample only contained 24 − x prolonged cell exposure data, and 24-hour washed sample contains no prolonged exposure, given that cell exposure was only allowed for the duration of a total time of 24 hours.

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