were given multiple doses of 3.75 mg/g /application PEGLA during D2 and killed on D6. Uteri were dissected out and fixed in Neutral Buffered Formalin before PEGLA immunoreactivity in the vagina and uterus was identified as previously described. Contraceptive efficacy of PEGLA. Mated female mice were given vaginal applications of PEGLA at two doses or PEGMSA control dissolved 1:2 in placebo gel during the peri-implantation period. The mouse vagina can hold only,15 ml, thus the dose of PEGLA able to be delivered per May 2011 | Volume 6 | Issue 5 | e19665 Contraceptive Action of Vaginal LIF Antagonist application was limited. Mated female mice were given a total of 5 applications at D2 9am, 3pm & 9pm and D3 9am & 3pm. It was not Talampanel cost possible to treat mice with a greater number of applications as this resulted in abortion in the SHAM treated mice. Treated mice were killed on D6 and the number of CL on the ovary and IS in the uterus counted. The effect of PEGLA on long-term fertility. Non-mated female mice were given IP injections of PEGLA or PEGylation reagent control at 12pm and 10pm and the following day at 10am. After 15 days the treated mice were paired until they plugged. Plugged mice were killed on D10 and the number of implantation sites in the uterus counted. The effect of PEGLA on bone remodelling. Mated and non-mated female mice were treated with PEGLA by IP injection or vaginal application. IP injections of PEGLA or PEGylation reagent control were given to mated and non-mated females at 12pm and 10pm and the following day at 10am. Vaginal applications of PEGLA or PEGMSA were given to mated females at D2 9am, 3pm & 9pm and D3 9671117 9am & 3pm). Treated mice were killed on D6 and tibiae analysed as previously described. Effect of PEGLA on EAE. In mice, LIF and CNTF prevent worsening of experimental auto-immune encephalomyelitis, an inducible animal model of multiple sclerosis. We investigated whether PEGLA could inhibit LIF action 
in the CNS and worsen EAE. EAE was induced in male mice using MOG 355 peptide. MOG 355 was delivered in equal parts peptide and Freund’s Complete Adjuvant containing Mycobacterium tuberculosis H37Ra. Each mouse received a total dose of 125 mg peptide, via 100 ml subcutaneous injections in each flank, as well as a subcutaneous injection of 50 ml in the base of the tail. On the initial day of EAE induction and D3, the mice also received an IP injection of 300 ng of Bordetella Pertussis toxin. Mice that reached EAE grade 2, were given water and powdered food in small Petri dishes at the base of the cage for easy access. Cotton wool buds were also added to cages for extra warmth. To monitor general health, mice were weighed every second day. Mice that were considered to be in poor health or that had lost their righting reflex were euthanized by administration of 100 ml lethabarb via IP injection. Disease severity was graded by a blinded observer using a standard EAE scoring system: 0 No clinical symptoms 1 Mild tail weakness 1.5 Mild tail weakness; mild gait abnormality 2 Complete tail atonia 2.25 Complete tail atonia; mild gait abnormality 2.5 Complete tail atonia; moderate gait abnormality 2.75 Complete tail atonia; paralysis of one hind limb 3 Complete tail atonia; paralysis of both hind limbs 3.5 Complete hind limb and tail paralysis plus animal unable to right when placed supine; animals were euthanized at this point 4 Death PEGLA Treatment. Mice received IP injections of PEGLA or PEG control every second day post
