B6 cg tg gfap cre 73.12mvs j mice

Adult Mus musculus males, females, and postnatal pups on a C57Bl/6J background were obtained from the Jackson Laboratory. B6.Cg-Tg(Gfap-cre)73.12Mvs/J mice (#012886, The Jackson Laboratory) used previously (Rothhammer et al., 2016 ) were crossed with B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J mice (#007909, The Jackson Laboratory) to generate Gfap(Cre/+);TdTomato(f/+) mice. Both male and female pups were sacrificed between P0-P3 for harvesting and culturing astrocytes and sex was assumed to be balanced and differences were not analyzed. Mice were kept in a pathogen-free facility at the Hale Building for Transformative Medicine at Brigham and Women’s Hospital in accordance with the IACUC guidelines, fed ad libitum on a 14/10-hour light/dark cycle. Mice were healthy and checked daily by veterinary staff. 8–10 week old mice were used for stereotactic injection and EAE induction. Mice were both drug and test naïve and not involved in previous procedures. All mice were at least doubly housed. All procedures were reviewed and approved under the IACUC guidelines at Brigham and Women’s Hospital.

To reduce HIF-1α levels in HSCs, HIF-1α^fl/fl mice (B6.129-Hif1a^tm3Rsjo/J, Jackson Laboratories, Bar Harbor, ME), described in detail previously (16 (link)), were crossed with mice expressing Cre recombinase under control of the glial fibrillary acidic protein (GFAP) promoter (B6.Cg-Tg(Gfapcre)73.12Mvs/J mice; Jackson Laboratories)(17 (link)). Both strains of mice were backcrossed to C57BL/6J for at least 12 generations. HIF-1α^fl/fl -GFAP Cre- (i.e., normal HIF-1α levels in GFAP-positive cells) and HIF-1α^fl/fl -GFAP Cre+ (i.e., decreased HIF-1α levels in GFAP-positive cells) littermates were used for these studies.
To monitor prolyl hydroxylase (i.e., EGLN) activity as an indirect measure of HIF activation, ODD-luc mice were used (F,FVB.129S6-Gt(ROSA)26Sor/J, Jackson Laboratories) (18 (link)).
All mice were maintained on a 12-h light/dark cycle under controlled temperature (18-21°C) and humidity. Food (Rodent Chow; Harlan-Teklad, Madison, WI) and tap water were allowed ad libitum. All procedures on animals were carried out in accordance with the Guide for the Care and Use of Laboratory Animals promulgated by the National Institutes of Health and approved by the Michigan State University IACUC.

Adult male and female mice and postnatal pups were used on a C57Bl/6J background (#000664, The Jackson Laboratory). B6.Cg-Tg(Gfap-cre)73.12Mvs/J mice^{52 (link)} (The Jackson Laboratory, #012886) mice were crossed with B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato) Hze/J mice^{53 (link)} (The Jackson Laboratory, #007909) to generate Gfap^cre/+TdTomato^f/+ (TdTomato^Gfap) mice. B6N.129-Rpl22tm1.1Psam/J mice^{54 (link)} (The Jackson Laboratory, #011029) were bred to Gfap^cre mice to generate Gfap^cre/+Ribotag^f/f (Ribotag^Gfap) mice. Aldh1l1^creERT2 mice^{55 (link)} (The Jackson Laboratory, #029655) were bred to Csf2rb^f/f mice^{39 (link)} to generate Aldh1l1^creERT2Csf2rb^f/f mice. Conditional deletion of Csf2rb was induced at 5 weeks of age with tamoxifen (225 mg/kg; Sigma-Aldrich, #T5648) diluted in corn oil (Sigma-Aldrich, #C8267); EAE was induced 3 weeks later. Mice were kept in a pathogen-free facility at the Hale Building for Transformative Medicine at Brigham and Women’s Hospital in accordance with the IACUC guidelines. Eight-to-twelve-week-old mice were used for stereotactic injection and EAE induction. Pups were killed between postnatal day 0 (P0) and P3 for collection and culture of astrocytes. All procedures were reviewed and approved under the IACUC guidelines at Brigham and Women’s Hospital.