Fig. 5. PFK158 initiates unfolded protein response in MPM cells.

a Fold changes in most of the UPR-related proteins after treatment with IC₅₀ of the PFK158 of each cell line at 24 h, followed by large-scale analyses of changes in protein level and their modification using reverse-phase protein arrays (RPPA) as described in the “Material and methods” section. One-way ANOVA was used to analyze the data. b ER and lysosomal activity were visualized by confocal microscopy stained with ER-tracker blue/white DPX and lysotracker red in H28 and EMMeso cell treated with PFK158 (10 μM) for different time point (0–6 h). c Graphical representation of the increase in ER-tracker blue and decrease in lysotracker red. MPM cells were loaded with Fura Red to stain intracellular Ca²⁺ pool and either observed under a fluorescent microscope d or analyzed by FACS e. Increase in ER stress marker proteins after pharmacological inhibition f and genetic knockdown g of PFKFB3 as demonstrated by Western blot