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. 2016 Sep 28;7(45):73292–73308. doi: 10.18632/oncotarget.12320

Figure 5. Only hyperphysiological citrulline can support regrowth of CRC spheroids after combined treatment with canavanine under arginine-deprived conditions.

Figure 5

(A) Monolayer HCT-116 and HT29 cells were exposed to –Arg medium containing 0.1 mM Cav with or without Cit supplementation for 3 days (grey bars). Afterwards, these specific media were exchanged to regular medium (RM), and the cells were allowed to grow for additional 3 days (white bars). Each data point presents the average number of viable cells (± SE) of three independent experiments. (B) Representative phase contrast images of HCT-116 and HT29 spheroids before treatment (day 0) and after 20 days of treatment with Cav in the specific media are shown. Scale bar = 200 μm. (C) The fractions of HCT-116 and HT29 spheroids that restored growth after 20 days of exposure to the combination treatment as described in (A) followed by an additional 20 days in RM post-treatment are shown. Each column represents data from at least 30 spheroids obtained in one experiment. n.d. – not detected.