POLR3 gene and protein expression dynamics in 4H 195 6 Subcellular localization of Pol III subunits To explore the functional implications of altered POLR3 gene expression in NES, we analyzed the subcellular localization of Pol IIIA and IIIB using immunocytochemistry (Supplementary Fig. 2). After removing one influential outlier, a 4H line with POLR3A variants, we did not observe a difference in subcellular localization of Pol IIIA subunit between control and 4H cells (χ2(1) = 2.161, p = 0.142; Fig. 3A). Similarly, no significant differences were found in Pol IIIB localization between controls and 4H cells ( χ2(1) = 0.300, p = 0.584; Fig. 3B). ICC analysis revealed modest contributions of individual differences to Pol IIIA (ICC = 0.096) and Pol IIIB (ICC = 0.1166) localization. These results do not show evidence for impaired localization caused by 4H variants and individual variability does not appear to be a major factor.
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