BALB/c mice (8‐week‐old) received a daily topical dose of 62.5 mg commercially available 5% IMQ cream (Aldara; 3M Pharmaceuticals, Tallinn, Estonia) on a shaved area on the back for 6 days. This dose was empirically determined to cause optimal and reproducible skin inflammation in mice. Two weeks before this model was established, we administered (i.v.) PBS or 5 μg hIL‐35‐Fc per mouse as prophylactic treatment every other day for a total of seven times, subsequent three more injections were conducted via the tail vein during establishment of the model.
Imq cream
Manufactured by 3M
Sourced in United States, United Kingdom
IMQ cream is a laboratory product manufactured by 3M. It is a sterile, white, water-miscible cream formulation.
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Lab products found in correlation
Invivomab rat igg2b isotype control, by BioXCell (1 mentions)
Control igg, by Proteintech (1 mentions)
Balb c mice, by Jackson ImmunoResearch (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Adhesive tape, by 3M (1 mentions)
Digimatic thickness gauge, by Mitutoyo (1 mentions)
C57bl 6 background, by Jackson ImmunoResearch (1 mentions)
C57bl 6, by Jackson ImmunoResearch (1 mentions)
30 protocols using imq cream
1
Prophylactic hIL-35-Fc Treatment in Imiquimod-Induced Skin Inflammation
2
Lupus Induction and pDC Depletion Model
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The lupus model was induced according to the previous reports with minor modifications (30 (link)). Briefly, the skin on the right ears of the mice was treated topically every other day, with 2 mg of 5% IMQ cream (Aldara, 3M Pharmaceuticals) for 6–12 consecutive weeks. Control mice were given the same dose of vehicle cream. Mice were treated with 10 mg/kg per day HCQ (Selleck Chemicals) by oral gavage during the induction period. For pDC depletion in vivo, mice were first injected intraperitoneally with 500 μg pure InVivoMAb anti-mouse antibody (Bio X Cell, BE0311) 4 days before IMQ application and then treated with antibodies every 4 days over the 8 weeks of topical treatment with IMQ. InVivoMAb rat IgG2b isotype control (Bio X Cell, BE0090) was used as the control.
3
Psoriasis Induction in TG2-/- Mice
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TG2−/− mice41 (link) were backcrossed 12 times to C57BL/6J. 10–11-week-old male mice were shaved and chemically depilated by Veet cream (Oxy-Reckitt Benckiser, Slough, UK) at 48 h before IMQ treatment. Daily topical dose (62.5 mg) of 5% IMQ cream (Aldara, 3 M Pharmaceuticals) was applied on the shaved back and the right ear for six consecutive days, as previously described24 (link). The psoriasis area and severity index (PASI) were scored by two independent researchers blind to genotype. Animal experiments were approved by the Seoul National University Institutional Animal Care and Use Committee.
4
Psoriasis Mice Model and Therapeutic Evaluation
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All of our experimental procedures and protocols were well approved by Zhejiang University of China in accordance with the Guide for Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH) (publication No. 96-01). Balb/c mice of wild type (6–8 weeks old, China Academy of Science, Shanghai, China) were selected for the animal experiments. To establish the mice model of psoriasis, mice were topically administered with imiquimod cream (IMQ, 3M, USA) on the back skin, which was used as a common psoriasis mice model.[16 (link)17 (link)] Briefly, IMQ cream of 5% was evenly applied to the alopecia area at a dose of 50 mg/cm2, once a day for 6 days. After the model was established, the mice were intraperitoneally injected with IgG control (Proteintech Co.), IL-17 antibody (Proteintech Co.), and IL-17+ HPSE inhibitor (OGT 2115). Seven and 10 days after treatments, the IMQ-treated area of the skin was isolated and subjected to immunohistochemistry (IHC) staining, ELISA, and RT-PCR analysis of HPSE and IL-17.
5
Induction and Treatment of Psoriasis-like Inflammation in Mice
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Eight- to 10-wk female BALB/c mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). Mice received a daily topical dose of 55 mg of IMQ cream (5%; Aldara; 3M Pharmaceuticals, London, Ontario, Canada) on the shaved areas of the mouse back and 5 mg on the right ear for 6 d to generate localized and generalized psoriasis. This dose was determined to induce psoriasis-like skin inflammation on day 3. On day 3, mice were divided into 5 groups, no IMQ treatment (normal), IMQ-treated (Pso.), IMQ plus either medium (Sham), 4 × 106 (link) non-IDO fibroblast (Pso. + Fib), or 4 × 106 (link) IDO-Red Cherry-expressing fibroblast injected (Pso. + IDOFib). Mice were intralesionally injected with either 4 × 106 (link) cells or control fibroblasts/100 μL of medium/spot on dorsal areas (4 spots/mouse).
The lesions were photographed every other day. The presence of erythema and scaling on the back and ears were scored on a scale from 0 to 4 by an unbiased observer (0, none; 1, slight; 2, moderate; 3, marked; and 4, very marked). Skin and ear thickness were measured every other day using the digital caliper. Upon euthanizing the mice on day 9, the size and weight of the auxiliary and inguinal lymph nodes and spleen were taken and evaluated.
The lesions were photographed every other day. The presence of erythema and scaling on the back and ears were scored on a scale from 0 to 4 by an unbiased observer (0, none; 1, slight; 2, moderate; 3, marked; and 4, very marked). Skin and ear thickness were measured every other day using the digital caliper. Upon euthanizing the mice on day 9, the size and weight of the auxiliary and inguinal lymph nodes and spleen were taken and evaluated.
6
Imiquimod-Induced Skin Inflammation Mouse Model
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Imiquimod (IMQ)-induced skin inflammation mouse model was established using IMQ cream.73 (link) Briefly, DMSO or Mdivi-1 treated mice were applied daily with IMQ cream (5%) (Aldara; 3M Pharmaceuticals) on the shaved back for 5 consecutive days. Mice were sacrificed on day 6. Skin cell suspensions from IMQ-treated skin were stained for CD45, CD11b, Gr-1, CD3, γδTCR and intracellular IL-17. Draining lymphocytes were also stained for CD3, γδTCR and intracellular IL-17 after PMA/I stimulation plus Golgi-Plug for 5h.
7
Exosome Treatment for Imiquimod-Induced Psoriasis
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All mice were randomly divided into 4 groups (𝑛= 6 mice/group): control group (Normal), IMQ group (IMQ), PBS-treated IMQ group (PBS + IMQ), and hucMSCs-Exo-treated IMQ group (hucMSCs-Exo + IMQ). All experimental procedures were conducted in accordance with the Animal Use and Care Committee of Shandong University. To establish the IMQ model, the C57BL/6 mice were treated with a daily topical dose (62.5 mg) of 5% IMQ cream (Aldara, 3M Health Care Limited) on the shaved dorsal skin for 6 days.
On days 0, 2 and 4 (Fig. 2(A)), the mice in the hucMSCs-Exo + IMQ group were subcutaneously injected with exosomes (50 µg/mouse), and the mice in the PBS + IMQ group were subcutaneously injected with PBS (50 µl/mouse). All mice were euthanized on day 7 for analysis. The dorsal skin of the mice was sectioned, and part of the skin was stained with haematoxylin and eosin (H&E) for histological evaluation. Another part was used for Western blotting.
On days 0, 2 and 4 (Fig. 2(A)), the mice in the hucMSCs-Exo + IMQ group were subcutaneously injected with exosomes (50 µg/mouse), and the mice in the PBS + IMQ group were subcutaneously injected with PBS (50 µl/mouse). All mice were euthanized on day 7 for analysis. The dorsal skin of the mice was sectioned, and part of the skin was stained with haematoxylin and eosin (H&E) for histological evaluation. Another part was used for Western blotting.
8
Psoriatic Dermatitis and K17 Inhibition
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Eight-week-old female BALB/c mice were purchased from the Department of Laboratory Animal Medicine of Fourth Military Medical University (Xi’an, China). All experimental procedures involving mice were performed in compliance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Review Committee for the Use of Animals of Fourth Military Medical University. For the generation of psoriatic dermatitis in mice, we applied a 5% IMQ cream (Aldara; 3M Health Care Ltd., Loughborough, UK) topically on both ears of the mice. To evaluate the effect of K17 inhibition on the development of psoriasis, 2.5 nmol K17 siRNA was mixed with 2 μL Lipofectamine 3000 (Invitrogen, Waltham, MA, USA) and 5 mmol/L emulsion matrix and applied topically to the left ear of the mice every day for 7 days after applying IMQ. The right ear of the mice was treated with NC siRNA.
9
Topical IMQ-Induced Psoriasis Model in BALB/c Mice
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All animals were housed in an animal room under temperature control (24–25 °C) and a 12:12 h light-dark cycle. Standard laboratory chow and tap water were available ad libitum. Experimental procedures were approved by the Mackay Memorial Hospital Review Board (New Taipei City, Taiwan) (project code: MMH-A-S-104-37, date: 1 January 2016–31 December 2018) and were performed in compliance with national animal welfare regulations (Council of Agriculture, R.O.C).
BALB/c mice (8-week-old females) were shaved on the back and psoriasis was induced by topical application of with IMQ cream, as described previously [17 (link)]. Accordingly, each mouse received a daily topical dose of 62.5 mg of commercially available IMQ cream (5%; Aldara; 3M Pharmaceuticals) on the shaved back, and 15 mg of IMQ on the right ear, for 6 consecutive days. Control mice (normal group) were treated with a control cream. For treatment, 25 µM C16 or dimethyl sulfoxide (DMSO as peptide vehicle) was mixed with the IMQ cream. Histologic analysis of back skin specimens was performed just after the termination of IMQ application (on day 7). Ear thickness was measured in duplicate using a digital micrometer before challenge and after IMQ treatment for 6 consecutive days.
BALB/c mice (8-week-old females) were shaved on the back and psoriasis was induced by topical application of with IMQ cream, as described previously [17 (link)]. Accordingly, each mouse received a daily topical dose of 62.5 mg of commercially available IMQ cream (5%; Aldara; 3M Pharmaceuticals) on the shaved back, and 15 mg of IMQ on the right ear, for 6 consecutive days. Control mice (normal group) were treated with a control cream. For treatment, 25 µM C16 or dimethyl sulfoxide (DMSO as peptide vehicle) was mixed with the IMQ cream. Histologic analysis of back skin specimens was performed just after the termination of IMQ application (on day 7). Ear thickness was measured in duplicate using a digital micrometer before challenge and after IMQ treatment for 6 consecutive days.
10
Mouse Model of Imiquimod-Induced Psoriasis
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Eight-week-old BABL/c mice were purchased from the Animal Department of Capital Medical University and were housed in a specific pathogen free facility. The housing conditions were controlled, with a temperature of 20–25 °C and a 12:12 hours light: dark cycle. Mice were numbered and randomly assigned to three groups, then, an area of 5 cm × 3 cm was shaved from the backs of all mice. Mice in the control group were given IMQ base cream topically and then intradermally injected with 100 μl normal saline, while mice in the normal saline (NS) group and C19 group received a daily topical application of 62.5 mg IMQ cream (5%, Aldara, 3 M Pharmaceuticals, Shanghai, China), followed by intradermal injection of 100 μl normal saline without or with 10 μl C19 peptide (Hybio Engineering, Shenzhen, China) for 6 consecutive days. At the end point of the experiment, all mice were killed by breaking the neck. Samples of skin from back lesions were collected for latter experiments.
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