Figure 1.

Exposure of prostate cancer cells to hypoxia causes dynamic changes in cell mechanical properties. (A) Immunoblot of Rb and α-tubulin protein expression in control shSCX and knockdown shRb LNCaP cells under hypoxic and normoxic conditions. Complete blot images are shown in Supplementary Information File 1, Figure S1. (B) PLOD2 mRNA accumulation in control shSCX and knockdown shRb LNCaP cells. Cells were maintained under normoxic conditions or 1% O₂ for 24 h. Gene expression was determined by quantitative real-time PCR after isolation and reverse transcription of total RNA. PLOD2 expression was normalized to constitutively active 36B4 gene expression. (C) Variation in measured NARBM (Normalized Amplitude of Resultant Bead Movement) of shRb and shSCX LNCaP cells after 24 h, 48 h and 72 h of exposure to hypoxia or normoxia. (D) Variation in shear modulus after 72 h of exposure to hypoxia or normoxia at different frequencies of applied force (0.1, 1, and 10 Hz). (E) Variation in viscosity of shRb and shSCX LNCaP cells after 72 h of exposure to hypoxia or normoxia. In each experiment, at least 10 cells were examined for each group and each experiment was repeated five times. Error bars represent ± S.D. *p < 0.05.