Physique S2. of experimental conditions needed for nutritional stress induction, in relation to the concentration of glucose UNC 0224 and glutamine in the medium, was combined with measuring cellular viability (Physique 2A), cellular proliferation (Physique 2B), and the amount of ROS generated at hour 48 (Physique 2C). In the beginning, the four nutritional conditions (NC1-NC4) were established, as explained in the Material and Methods section, to which only the malignancy cells were uncovered. Then, Tnfrsf1b based on the data obtained, this research was extended to IMR-90, through applying nutritional conditions NC2 and NC3. NC1 should be considered the control condition. Open in a separate window Physique 2 The viability, proliferation and generation of ROS in malignancy cell lines (ACC) and IMR-90 (DCF) after exposure to NC1-NC4 and NC1-NC3, respectively, for 48 hours. One-way ANOVA with Tukey post-hoc test was used to test the differences with regard to nutrient conditions. The values are shown as the mean 95% CI. N = 3. * < 0.05; ** < 0.01; *** < 0.001; **** < 0.0001. As offered in Physique 2A, the viability of the malignancy cell lines, UNC 0224 regardless of their genetic background, was similar under the given conditions. Predictably, the most rigorous decrease in cellular viability (up to 70%) was recorded for all those three malignancy cell lines in the medium without glucose and glutamine (NC3), when compared to both NC1 and NC2 (< 0.0001). The presence of glutamine in a medium without glucose (NC4) was beneficial for the viability of all malignancy cell lines (Physique 2A). It was also beneficial for the cellular proliferative capacity (Physique 2B) of Detroit 562 (= 0.0007) and Cal 27 (< 0.0001), but not FaDu. The strongest ROS generation was associated with condition NC3. The presence of glutamine in a medium without glucose (NC4) led to a decreased ROS generation in all three malignancy cell lines (Physique 2C). Under NC2, the generation of ROS in Cal UNC 0224 27 and FaDu was stronger (= 0.0055 and < 0.0001, respectively) than in Detroit 562 (Figure 2C). When examined, these data indicated that this malignancy cell lines showed some interesting and unique features. Under NC2, the generation of ROS was not significantly increased only in Detroit 562. Under NC4, when there is a lack of glucose, FaDu was far less sensitive to the rescuing effect of glutamine on proliferative capacity. These data are also very indicative regarding the degree of cellular sensitivity to glutamine deprivation, showing that Cal 27 and Detroit 562 were more dependent on glutamine than was FaDu. Knowing that non-transformed cells are highly dependent on glucose, and relying on the data obtained with the UNC 0224 malignancy cell lines (Physique 2ACC), we chose to continue the experiments using the mildest (NC2) and the most strong condition (NC3), now including the IMR-90 fibroblasts. They were considered a good control system, to compare with the malignancy cell lines. The Physique 2DCF represent IMR-90 response to NC2 and NC3. The viability of IMR-90 was unique in the extreme sensitivity of this cell line to the moderate glucose deprivation, during 48 hours (NC2; < 0.0001) (Physique 2D). The viability and proliferative capacity after 48 hours in NC2 (Determine 2D,E) seems to be a maximal effect of a critical nutrient deprivation (glucose) because 48 h of cultivation in the medium without glucose and glutamine (NC3) did not influence these cellular parameters further. However, the generation of ROS did differ between NC2 and NC3 (= 0.0019) (Figure 2F), although not as strong as in the cancer cell lines (< 0.0001) (Physique 2C). The lack of a significant.