Extended Data Fig. 5: Hypoxia-induced macropinocytosis depends on oncogenic KRAS in PDACs.
(a) Immunoblot analysis of FLAG and T7, showing inducible expression of stably transduced FLAG-tagged KRASG12V BxPC-3 cells and T7-tagged KRASG12V HeLa cells after addition of doxycycline (1 μg/mL for 2 days). ACTB or vinculin were used as a loading control. (b-e) Representative images (b, d) and quantification (c, e) of DQ-BSA fluorescence (green) in PANC-1 (b, c) and MIA PaCa-2 (d, e) cells under normoxia (21% O2) and hypoxia (0.5% O2). Nuclei are labeled with DAPI (blue). Scale bar, 10 μm. In c, e, data are presented relative to normoxia. c, e, Bars represent mean ± s.e.m. At least 500 (c, d) cells were quantified in each biological replicate (n = 3). Statistical significance was determined by two-tailed unpaired t-test.
