5.
mRNA perturbation of Ipl1 leads to amelioration of growth suppression and restores H3S10ph levels in (PR)50 overexpression yeast. (A) Serial growth assay verifying toxicity elicited by (PR)50 overexpression in BY4741 (Parental) and Ipl1 DAmP yeast. (B) Western blot verifying presence of (PR)50 in parental and Ipl1 DAmP (PR)50 yeast. (C) Confocal microscopy of parental and Ipl1 DAmP control and (PR)50 yeast stained with α-FLAG (red) and a DAPI (blue) costain. (D) Column scatterplot depicts the number of aggregates per cell from either parental control (orange) or (PR)50 (blue) yeast. Each point represents a cell from each group. (n = 149–152), **** = p ≤ 0.0001. (E) Column scatterplot depicts the number of aggregates per cell from either Ipl1 DAmP control (orange) or (PR)50 (blue) yeast. Each point represents a cell from each group. (n = 102–111), **** = p ≤ 0.0001. (F) Western blot of parental and Ipl1 DAmP control and (PR)50 yeast probing for H3S10ph and Histone H3 Total. (G) Column scatterplot quantifies the relative density of H3S10ph levels in parental control and (PR)50 yeast. Each point in the graph represents a separate experiment with a different biological replicate. (n = 5; *** = p ≤ 0.001). (H) Column scatterplot quantifies relative density of H3S10ph levels in Ipl1 DAmP control and (PR)50 yeast. Each point in the graph represents a separate experiment with a different biological replicate. (n = 5; *** = p ≤ 0.001).