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Alternatively, for mice receiving a dose of 1 × 10 CFU, the original bacterial stock was diluted in PBS to 1 × 10 CFU/mL, and 100 μL was injected per mouse as above.
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C57BL/6J mice as above were assigned to vehicle, Boc5 or Boc5 plus exendin(9 39) treatment groups (n≥8).
In intact hpg mice, as above, the ARC kisspeptin immunostaining pattern again showed dark somatic staining with reductions in fiber staining, with no changes in pattern with OVX (Fig. 7C).
CD11c+ DCs were isolated from adult and old mice as above, and graded numbers were seeded in 100 μL RPMI 1640 + 10% FCS in 96-well U-bottom plates.
Hepatocytes (5 × 10) isolated from WT or Idh1 KO mice as above were lysed in the NADP Extraction Buffer supplied with the Fluoro NADP/NADPH Detection Kit (Cell Technology, Fremont, CA, USA).
For the anti-tumor efficacy experiment, tumors were grown in nude mice as above.
For secondary transplants, bone marrow cells (105 total human CD19+ cells) were injected by tail vein into irradiated NOD/Scid mice as above.
For antigen-specific MLR, 2.5×105/ml irradiated (2000 rad) Jak2−/− BMDCs or splenic DCs were pulsed with 1 µM OVA peptide and then co-cultured with 5×105/ml splenocytes isolated from OT-1 transgenic mice as above.
In the second study, stably transfected TRAMP-C2 cells were collected, irradiated (10 000 rads), and injected into mice as above.
For quantification of soluble huntingtin levels in rilmenidine-treated mouse brains, protein was extracted from seven placebo-treated mice and six rilmenidine-treated mice as above.
To further address the impact of ER stress on renal fibrosis, we induced renal fibrosis by UUO in Chop-deficient and wild-type (WT) mice as above.
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