Mice (8-12 weeks old) were infected naturally with the murine filaria Litomosoides sigmodontis14 (link),15 (link) and subsequently co-infected with Herpes Simplex Virus 2 (HSV-2; 5x105 PFU intravaginally, strain G, ATCC VR-734) for 7 days.16 (link) To facilitate HSV-2 infection and synchronize the estrous cycle, progesterone (2 mg/19 g body weight) (Depo-Provera®, Pfizer, New York, NY, USA) was administered subcutaneously 7 days prior to HSV-2 infection. The results shown here are representative of three experiments, each with at least four mice.
Depo provera
Manufactured by Pfizer
Sourced in United States, Belgium
Depo-Provera is a type of injectable contraceptive medication. It contains the progestin hormone medroxyprogesterone acetate, which is used to prevent pregnancy.
Automatically generated - may contain errors
Lab products found in correlation
Jadelle, by Bayer (2 mentions)
C57bl 6 mice, by Charles River Laboratories (2 mentions)
Microfiber swab, by Thermo Fisher Scientific (2 mentions)
Chlamydia isolation medium, by Trinity Biotech (2 mentions)
Cpg odn 1826, by InvivoGen (1 mentions)
17β estradiol, by Merck Group (1 mentions)
Xylazine hydrochloride, by Bayer (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Imagexpress pico automated cell imaging system, by Molecular Devices (1 mentions)
Ilium ketamil, by Troy Laboratories (1 mentions)
Ilium xylazil 20, by Troy Laboratories (1 mentions)
Lethabarb, by Virbac (1 mentions)
Balb cbyj mice, by Charles River Laboratories (1 mentions)
Rnalater solution, by Thermo Fisher Scientific (1 mentions)
Mirena iud, by Bayer (1 mentions)
Female c57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Xylazine, by Bayer (1 mentions)
Living image software, by PerkinElmer (1 mentions)
Vicryl 5 0, by Johnson & Johnson (1 mentions)
Chirocaine, by Abbvie (1 mentions)
55 protocols using depo provera
1
Litomosoides and HSV-2 Co-Infection in Mice
2
Murine Chlamydia Infection Model
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Female mice (6‐8 weeks) were hormonally synchronized with 2.5 mg depot medroxyprogesterone (Depo Provera, Pfizer, New York, USA) subcutaneously 1 week prior to intravaginal infection with 5 × 104 inclusion forming units (IFUs) of C. muridarum in 10 μl volume.
3
Chlamydia Infection in Knockout Mice
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All mouse lines were maintained at animal facilities at Duke University Medical Center. Animal protocols were approved by the Institutional Animal Care and Use Committees at Duke University. Wild-type C57BL/6 (Jackson Laboratories), Irgm2−/−, pan-Irgm−/− (52 (link)), Irgm3−/−, Irgm1/m3−/− (33 (link)), and Rag1/pan-Irgm−/− mice were used. All knockout mouse lines are on a C57BL/6 background as described in the relevant citations. For all experiments, 6–12-week-old mice were injected subcutaneously with 2.5 mg medroxyprogesterone acetate (Depo-Provera, Pfizer) diluted in sterile PBS 1 week prior to infection. Chlamydia EBs were diluted in sucrose-phosphate-glutamate (SPG) buffer (10 mM sodium phosphate, 220 mM sucrose, and 0.50 mM L-glutamic acid). For transcervical infection, 5 µL of diluted Chlamydia was instilled directly into the uterus using a Non-Surgical Embryo Transfer device (Braintree Scientific). For in vivo C. trachomatis infection, 5 × 106 IFU was delivered per mouse, and for in vivo C. muridarum infection, 2.5 × 105 IFU was delivered per mouse. For all mock infections, sterile SPG was used. Infected mice were monitored over the course of each infection.
4
Depo-Provera Enhances Mucosal Immunity
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150 days post MA immunization, mice were injected subcutaneously with 3 mg of Depo-Provera (Pfizer), a long-lasting progestin, in 100 μL of sterile PBS 5 days before intravaginal challenge. Groups of mice were challenged by depositing 20 μg of HIV-1 CN54 gag308–318 peptide (ProImmune) and 30μg of CpG ODN 1826 (InvivoGen) into vaginal wall and monitored for 5 days.
5
Modulation of Embryo Implantation in Mice
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Pseudopregnant mice were ovariectomized in the evening of D3, and siliconized medroxyprogesterone acetate (Depo Provera, 1.0 mg/mouse; Pfizer, Inc.; NY, USA) was subcutaneously injected to induce delayed implantation. Three days later, the following treatments were performed either to the mice; 3 ng or 20 ng of 17β-estradiol (Sigma-Aldrich) by subcutaneous injection, 25 μg of rLIF protein, or 200 μg of rFGF2 or rFGF9 protein by intraperitoneal injection, recombinant IGF1 protein (Somazon; 100 μg/mouse in every 6 hr for 24 hr; Orphan Pacific, Tokyo, Japan) by subcutaneous injection, or vehicle (corn oil or phosphate buffered saline). The uteri were collected 24 hr after first injection in order to analyze cell proliferation.
6
Fc-Fused IL-7 Enhances Mucosal Absorption
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Example 2
The codon-optimized human IL-7 and granulocyte colony-stimulating factor (G-CSF) genes were individually fused with a hybrid Fc-fragment. The schematic structure of Fc-fused IL-7 is shown in
3 mg of medroxyprogesterone acetate (Depo-Provera, Pfizer) was subcutaneously injected to mice in a diestrus state 4 days before treatment. The mice were anesthetized by intraperitoneal injection with 100 mg/kg ketamine (Yuhan) and 10 mg/kg xylazine hydrochloride (Bayer) in PBS. Then, 10 μg of rIL-7, IL-7-Fc or G-CSF-Fc were mixed with PBS and applied (administered) on the vaginal mucosal tissues using a micropipette.
PBS, rIL-7 and IL-7-Fc were intravaginally administered to mice (n=7/group), and serum concentration of IL-7 was measured by human IL-7 ELISA. The results are shown in
These results reveal that the application of the Fc-fused protein on the mucosal epithelium enables genital-epithelial barrier transcytosis.
7
Chlamydia Challenge Protocol for Mice
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All animals received a subcutaneous injection of 2.5 mg of medroxyprogesterone acetate (Depo-Provera, Pfizer) one week prior to challenge. Three weeks after the final vaccination, mice were challenged intra-vaginally with 107 IFU of Ct serovar D/UW3/Cx EBs as previously described [7] (link), [17] (link). Cervico-vaginal swabs were obtained on days 1, 3, 7, 14 and 22 post-challenge, and the total number of inclusions per well were enumerated in a blinded fashion using fluorescence microscopy.
8
Synchronizing Mouse Estrous Cycle for Chlamydia Infection
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Ten and three days before C.t. SvA or SvIa challenge, the oestrous cycle was synchronized by injection of 2.5 mg of medroxyprogesteronacetat (Depo-Provera, Pfizer, Ballerup, Denmark), increasing mouse susceptibility to chlamydial infection by prolonging dioestrus. The mice were challenged i.vag. with 1 × 106 C.t. SvA/HAR-13 or SvIa in 10 µl of SPG buffer. At different time points post infection (PID3, 7, 10, 14, and 17), the mice were swabbed. Swabs were vortexed with glass beads in 0.6 ml of SPG buffer and stored at –80 °C until analysis. The infectious load was assessed by infecting 48-plate wells seeded with McCoy cells with the swab material undiluted and twofold diluted. Inclusions were visualized by staining with polyclonal rabbit anti-MOMP serum made in our lab, followed by an Alexa 488-conjugated goat anti-rabbit immunoglobulin (1:500–1:1000, Invitrogen #A11008). Background staining was done with Propidium iodide (Invitrogen). IFU was enumerated by fluorescence microscopy either manually or by using an automated cell imaging system (ImageXpress Pico automated Cell imaging system) (Molecular Devices) counting 50% of each well. If no IFU were detected in the counted area, 100% of each well was counted manually. Culture-negative mice were assigned the limit of detection of 4 IFU/mouse representing one IFU in the tested swab material (1/4 of the total swab material).
9
Chlamydia muridarum Infection Model
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Mice were allocated into groups using a minimization strategy to balance variables, namely age and weight, across groups. Age matched, adult (6–8 weeks old), female WT, Ifne-/-, or Il15-/- C57BL/6 mice were pre-treated with 2.5 mg depot medroxyprogesterone acetate (Depo-Provera; Pfizer, NY, USA) subcutaneously to prime for infection and synchronize their estrogenic cycles. Seven days later, mice were infected by intravaginal inoculation with 5 × 104 IFU C. muridarum (ATCC VR-123) in 10 μL sucrose-phosphate-glutamate buffer (SPG; 10 mM sodium phosphate [pH 7.2], 0.25 M sucrose, 5mM L-glutamic acid) or sham-infected with SPG alone under ketamine:xylazine anesthesia (80 mg/kg:5 mg/kg IP; Ilium Ketamil® and Ilium Xylazil-20®; Troy Laboratories, Glendenning, Australia), as described previously (Asquith et al, 2011 (link); Fung et al, 2013 (link); Fig. EV4A ). Mice were sacrificed by sodium pentobarbital (250 mg/kg IP; Lethabarb; Virbac, Milperra, NSW, Australia) overdose at 3dpi for characterization of immune responses or 14dpi for quantification of FRT pathology.
10
Murine Model of Cervical HPV Infection
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