Cell Stem Cell, Volume 8 Supplemental Information LRRK2 Mutant iPSC-Derived DA Neurons Demonstrate Increased Susceptibility to Oxidative Stress Ha Nam Nguyen, Blake Byers, Branden Cord, Aleksandr Shcheglovitov, James Byrne, Prachi Gujar, Kehkooi Kee, Birgitt Schüle, Ricardo E. Dolmetsch, William Langston, Theo D. Palmer, and Renee Reijo Pera Inventory of Supplemental Information 1. Supplemental Figures and Legends Figure S1, related to Figure 1 Figure S2, related to Figure 2 Figure S3, related to Figure 4 Figure S4, related to Figure 5 Figure S5, related to Figure 6 2. Table S1, related to Figure 3 3. Table S2 (Cell Counts), related to Figure 5 4. Table S3 (Cell Counts), related to Figure 6 5. Table S4 (Cell Counts), related to Figure 7 Supplemental Tables S2, S3 and S4 are provided in MS Excel spreadsheets. 6. Movie S1 (Beating Cardiomyocytes), related to Figure 1 7. Supplemental Experimental Procedures 8. Supplemental References
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Cell Stem Cell, Volume 8
Supplemental Information
LRRK2 Mutant iPSC-Derived DA Neurons
Demonstrate Increased Susceptibility
to Oxidative Stress Ha Nam Nguyen, Blake Byers, Branden Cord, Aleksandr Shcheglovitov, James Byrne, Prachi Gujar, Kehkooi Kee, Birgitt Schüle, Ricardo E. Dolmetsch, William Langston, Theo D. Palmer, and Renee Reijo Pera
Inventory of Supplemental Information
1. Supplemental Figures and Legends
Figure S1, related to Figure 1
Figure S2, related to Figure 2
Figure S3, related to Figure 4
Figure S4, related to Figure 5
Figure S5, related to Figure 6
2. Table S1, related to Figure 3
3. Table S2 (Cell Counts), related to Figure 5
4. Table S3 (Cell Counts), related to Figure 6
5. Table S4 (Cell Counts), related to Figure 7
Supplemental Tables S2, S3 and S4 are provided in MS Excel spreadsheets.
6. Movie S1 (Beating Cardiomyocytes), related to Figure 1
7.25 – intracellular. Correspondingly, spontaneous postsynaptic currents, measured under this experimental
condition at -70 mV, are a superposition of EPSCs and IPSCs (ECl- ~ –46 mV). Data were collected and initially
analyzed with the Patchmaster software (HEKA). Further analysis and statistics were performed using
IgorPRO and MS Excel.
Karyotyping and Teratoma Assay
Spectral karyotyping (SKY) was as described (Nguyen and Reijo Pera, 2008). For teratoma formation, iPSCs
from a confluent 10-cm dish were harvested by Collagenase IV treatment and injected subcutaneously into
female SCID mice (BL17SCID, Charles River). After 5-8 weeks post-transplantation, tumors were dissected
from the host, fixed in 4% paraformaldehyde/PBS solution, paraffin embedded, sectioned and stained with
Hematoxylin and Eosin.
REFERENCES Chambers, S.M., Fasano, C.A., Papapetrou, E.P., Tomishima, M., Sadelain, M., and Studer, L. (2009). Highly efficient neural conversion of human es and ips cells by dual inhibition of smad signaling. Nat. Biotechnol. 27, 275-280.
Eisen, M.B., Spellman, P.T., Brown, P.O., and Botstein, D. (1998). Cluster analysis and display of genome-wide expression patterns. Proc. Natl. Acad. Sci. 95, 14863-14868.
Nguyen, H.N., and Reijo Pera, R.A. (2008). Metaphase spreads and spectral karyotyping of human embryonic stem cells. CSH Protocols doi:10.1101/pdb.prot5047.