Figure 3. Hypoxia induces lipid accumulation and promotes proliferation following reoxygenation in prostate cancer LNCaP cells.

LNCaP cells were seeded at 50,000 cells/well in a 6 well plate. After 36 hrs, media was changed and cells were either transferred to hypoxia chamber (1% O₂) or continued under normoxia (21% O₂). After 48 hrs, one set of hypoxic plates was transferred to normoxia and maintained in normoxic conditions for 24, 48 and 72 hrs, while another set remained under hypoxia. A.–B. After completing the respective time periods, cells were fixed with 10% formaldehyde and stained with ORO. Representative pictures (at 200X) for ORO staining are shown. Lipid content quantitation was carried out by dye elution using isopropanol, and absorbance was measured by spectrophotometer at 500 nm. C.–D. At every time-point, total cell number and dead cell percentage were also measured. *, p ≤ 0.001; #, p ≤ 0.01; $, p ≤ 0.05.