Nox1 KO, Mapk14f/f, and Mapk7f/f conditional KO mice were generated as described previously (Nishida et al, 2004 (link); Matsuno et al, 2005 (link); Wang et al, 2006 (link)). These mice were crossed with transgenic Gt (ROSA)26Sortm1(EYFP)Cos mice (designated R26R-Eyfp; a gift from Dr. F Costantini, Columbia University Medical Center, New York, NY) to introduce a donor cell marker for transplantation experiments. GS cells from a transgenic mouse line of B6-TgR(ROSA26)26Sor mice (designated ROSA) (Jackson Laboratory) were previously described (Kanatsu-Shinohara et al, 2003 (link)), and GS cells were derived from Mapk14f/f and Mapk7f/f conditional KO mice using culture medium based on Iscove’s modified MEM (Invitrogen), which was supplemented with 10 ng/ml human FGF2, 15 ng/ml recombinant rat GDNF (both from Peprotech), and 1% FBS, as described previously (Kanatsu-Shinohara et al, 2014 (link)). Cultures were maintained on mitomycin C–treated mouse embryonic fibroblasts (MEFs). Where indicated, hydrogen peroxide (30 μM) or NAC (0.5 mM; Sigma) was added to the cultures. The chemicals used in the study are listed in Table S7. FGF2 and GDNF are supplemented in the medium unless stated otherwise.
Table S7