Ted a spherical 3D cell model with a diameter of 50 mm, which is divided into cubic Pleuromutilin custom synthesis compartments with identical edge length of 1.52 mm to allow reaction-diffusion simulations in 3D space (Figure 2A). The compartments were divided into three3D Spatial Effect on Nuclear NF-kB Avasimibe site OscillationFigure 1. Schematic view of the temporal model and its simulation result. (A) The model includes IKK activation, subsequent phosphorylation and proteosomal degradation of inhibitory protein IkBa, IkBb, and IkBe, activation of NF-kB, and its translocation to nucleus where a gene for IkBa is expressed in the NF-kB-dependent manner. (B) The simulated oscillation of the temporal model (red line) and an experimental observation by Sung, M.L. et al., PLos ONE, 2009 [25] (dots) are shown. The concentration of nuclear NF-kB (NF-kBn) is normalized to the maximum value. doi:10.1371/journal.pone.0046911.gthe control conditions, f, tfp, tp, and td are 0.139 mHz, 0.617 hrs, 9.32 hrs, and 7.14 hrs, respectively.N/C ratio alters the oscillation patternIt is reported that in human cancer patients, both nuclear volume and N/C ratio are increased [52,55], and more importantly, they are positively correlated with the progression and malignancy of the cancer [56,57,58,59]. Hence, it isimportant to determine if the oscillation pattern changes with N/C ratio changes. We summarized all oscillations tested for N/C ratios from 2.9 to 19 along a time from 0 to 10 hrs with amplitudes in red and blue for higher and lower NF-kBn, respectively, together with ordinary plots of time courses at N/C ratios of 2.9, 8.3 (control), and 19 (Figure 3A). This representation tells us overall alteration of oscillation pattern by changes in N/C ratio. It is clearly seen thatFigure 2. 3D model requires a different parameter set from that used in the temporal model. (A) 3D model of spherical cell with diameter of 50 mm, which is divided into compartments enabling reaction-diffusion simulation. Red compartments indicate the nuclear membrane compartments. (B) Middle panel is the 3D simulation result with the same reaction rate constants as in the temporal model. The simulation result shows much lower oscillation frequency as compared to the temporal model shown in the top panel. Bottom panel is the oscillation in the 3D simulation with modified reaction rate constants. (C) No 
combination of diffusion coefficient and the location of IkBs protein synthesis (blue plane) gives comparable oscillation frequency as in the temporal model (orange plane). The range of D is 10213 to 10210 m2/s with three locations of IkBs protein synthesis, which are indicated by three icons. (D) We defined oscillation frequency f, height of the first peak A0, time to the first peak tfp, decay time constant of the peak tp, and decay time constant td of successive amplitudes A0, A1, A2…., as parameters characterizing nuclear NF-kB oscillation. doi:10.1371/journal.pone.0046911.g3D Spatial Effect on Nuclear NF-kB Oscillationthe oscillation frequency remains largely unchanged by changes in N/C ratio because the intervals of the color changes along the horizontal axis are almost the same for all N/C values tested. This is also shown by Fourier analysis (Figure 3B). There is no significant change in tfp, either because the time to the first peak (reddish, yellowish or greenish color depending on N/C ratio) does not change much in Figure 3A and is quantitatively shown by the lack of change in tfp (Figure 3D). However, there is a lar.Ted a spherical 3D cell model with a diameter of 50 mm, which is divided into cubic compartments with identical edge length of 1.52 mm to allow reaction-diffusion simulations in 3D space (Figure 2A). The compartments were divided into three3D Spatial Effect on Nuclear NF-kB OscillationFigure 1. Schematic view of the temporal model and its simulation result. (A) The model includes IKK activation, subsequent phosphorylation and proteosomal degradation of inhibitory protein IkBa, IkBb, and IkBe, activation of NF-kB, and its translocation to nucleus where a gene for IkBa is expressed in the NF-kB-dependent manner. (B) The simulated oscillation of the temporal model (red line) and an 
experimental observation by Sung, M.L. et al., PLos ONE, 2009 [25] (dots) are shown. The concentration of nuclear NF-kB (NF-kBn) is normalized to the maximum value. doi:10.1371/journal.pone.0046911.gthe control conditions, f, tfp, tp, and td are 0.139 mHz, 0.617 hrs, 9.32 hrs, and 7.14 hrs, respectively.N/C ratio alters the oscillation patternIt is reported that in human cancer patients, both nuclear volume and N/C ratio are increased [52,55], and more importantly, they are positively correlated with the progression and malignancy of the cancer [56,57,58,59]. Hence, it isimportant to determine if the oscillation pattern changes with N/C ratio changes. We summarized all oscillations tested for N/C ratios from 2.9 to 19 along a time from 0 to 10 hrs with amplitudes in red and blue for higher and lower NF-kBn, respectively, together with ordinary plots of time courses at N/C ratios of 2.9, 8.3 (control), and 19 (Figure 3A). This representation tells us overall alteration of oscillation pattern by changes in N/C ratio. It is clearly seen thatFigure 2. 3D model requires a different parameter set from that used in the temporal model. (A) 3D model of spherical cell with diameter of 50 mm, which is divided into compartments enabling reaction-diffusion simulation. Red compartments indicate the nuclear membrane compartments. (B) Middle panel is the 3D simulation result with the same reaction rate constants as in the temporal model. The simulation result shows much lower oscillation frequency as compared to the temporal model shown in the top panel. Bottom panel is the oscillation in the 3D simulation with modified reaction rate constants. (C) No combination of diffusion coefficient and the location of IkBs protein synthesis (blue plane) gives comparable oscillation frequency as in the temporal model (orange plane). The range of D is 10213 to 10210 m2/s with three locations of IkBs protein synthesis, which are indicated by three icons. (D) We defined oscillation frequency f, height of the first peak A0, time to the first peak tfp, decay time constant of the peak tp, and decay time constant td of successive amplitudes A0, A1, A2…., as parameters characterizing nuclear NF-kB oscillation. doi:10.1371/journal.pone.0046911.g3D Spatial Effect on Nuclear NF-kB Oscillationthe oscillation frequency remains largely unchanged by changes in N/C ratio because the intervals of the color changes along the horizontal axis are almost the same for all N/C values tested. This is also shown by Fourier analysis (Figure 3B). There is no significant change in tfp, either because the time to the first peak (reddish, yellowish or greenish color depending on N/C ratio) does not change much in Figure 3A and is quantitatively shown by the lack of change in tfp (Figure 3D). However, there is a lar.
