Skip to main content
. 2017 Dec 1;37(2):201–218. doi: 10.15252/embj.201797630

Figure 6. In vitro characterization of later passage ICL MEFs.

Figure 6

  1. Growth curve of the later passage ICL lines Ch10 and Ch14 under adherent in vitro culture conditions (n = 3 for each data set and error bars denote SD, **P < 0.005 for Ch10 and ***P < 0.0001 for Ch14). See also Appendix Fig S3A. Two‐tailed unpaired t‐test was used to determine statistical significance.
  2. Colony formation in ICL and control MEFs. 5,000 cells were seeded in 10‐cm plates and stained with methylene blue after 3 days. Representative colony formation images in Ch10 and Ch14 are shown (left) and quantification of colonies formed (right) in the 4 ICL lines (n = 3 for each data set and error bars denote SD, **P < 0.005 for Ch10, Ch12, and Ch14 and ns for Ch9). Two‐tailed unpaired t‐test was used to determine statistical significance.
  3. Quantification of growth in ultra‐low adherent, sphere‐forming conditions for the four ICL lines (n = 3 for Ch10 and Ch14, n = 6 for Ch9 and Ch12 and error bars denote SD, ***P < 0.0001 for Ch9, **P < 0.001 for Ch10 and *P < 0.05 for Ch12 and Ch14). Two‐tailed unpaired t‐test was used to determine statistical significance.
  4. Analysis of metabolites (glucose consumed, lactate produced and glutamine consumed) for the ICL lines. Spent media was analyzed after each ICL, and control line was grown in culture for 3 days. Media in identical culture conditions, but with no plated cells, was used as a baseline for all samples (n = 3 for each data set and error bars denote SD; Glucose consumed: ***P < 0.0005 for Ch9 and Ch14, ***P = 0.0005 for Ch10 and *P < 0.05 for Ch12; Lactate produced: ***P < 0.0005 for Ch9 and Ch12, ***P = 0.0008 for Ch10, and ***P = 0.0005 for Ch14; Glutamine consumed: ***P < 0.0005 for Ch9 and Ch14). Two‐tailed unpaired t‐test was used to determine statistical significance.