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. 2020 Feb 25;30(8):2729–2742.e4. doi: 10.1016/j.celrep.2020.01.080

Figure 2.

Figure 2

PTEN Loss in KRAS-Driven PDAC Tumor Cells Upregulates Macropinocytosis

(A) Macropinocytosis assay using TMR-dextran as a marker of macropinosomes (red staining) in KPC and KCPTEN cells. Nuclei stained with DAPI (blue) (4′,6-diamidino-2-phenylindole) and Alexa 488 phalloidin (green) for actin staining (cellular periphery). The amiloride macropinocytosis inhibitor EIPA was used as a control at 50 μM.

(B) Quantification of TMR-dextran fluorescence. An average of 30 z stack images was acquired per condition. Values were normalized to the average of the vehicle-treated (DMSO) control.

(C) Lysosomal processing of extracellular protein uptake via macropinocytosis was assessed by DQ-BSA fluorescence.

(D) Quantification of DQ-BSA fluorescence. An average of 50 z stack images were acquired per cell line. Values were normalized to the average value of KPC1.

For (B) and (D), error bar represents SEM of two biological experiments, each conducted with three technical replicates. Scale bar represents 20 μm. Significance was determined by one-way ANOVA. ns, non-significant. ∗∗∗∗p < 0.0001 and ∗∗∗p < 0.001.