JCS03004 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Analysis of (A, B) osteogenic and (C, D) adipogenic differentiation of MSCs. (A) Alkaline phosphatase activity of MSCs cultured in standard conditions and (B) after 10 days of osteogenic induction treatment. (C) No lipid droplet formation was seen in MSCs cultured in control medium. (D) Oil Red O staining showing lipid droplets after 21 days of adipogenic induction (objective, 20×).

• Supplemental Figure 2 -

  Fig. S2. Effect of serum on MSC survival. (A) Photomicrographs (objective, 40×) of MSCs with 20% FCS in control conditions and (B) after 72 hours of CoCl₂ treatment. (C) Serum deprivation induced MSC death in control conditions and (D) after 72 hours of CoCl₂ treatment.

• Supplemental Figure 3 -

  Fig. S3. Effect of desferroxamine (DFX) on MSCs. (A) MSC morphological changes observed after 3 and 5 days of 150 μM DFX treatment by phase-contrast microscopy (objective 40×). (B) Real-time RT-PCR analyses of neuronal genes and osteogenic marker genes after 6 and 24 hours of DFX treatment. The values are represented as fold change compared to control ± s.e.m., n=3, *P<0.05, univariate Student’s t-test. (C) HIF-1α immunostaining observed on MSCs with confocal microscopy after 3 hours of DFX treatment. Bar, 20 μm.

• Supplemental Figure 4 -

  Fig. S4. Clonal analysis of MSC. (A,B,C) Single cells were isolated after diluting serially MSC in 96-well plates (objective 40×). (D,E,F) These cells were expanded for 12 days in standard culture medium and (G, H, I) induced with CoCl₂/Y-27632 treatment for 3 days (objective 20×).