Journal: International Journal of Medical Sciences

Article Title: Bombesin receptor-activated protein homolog deficiency altered the pattern of pathological changes of psoriasis - like skin lesion in mice

doi: 10.7150/ijms.89492

Figure Lengend Snippet: TSLP expression was increased by down-regulation of BRAP or its mouse homolog. (A) Double immunofluorescence staining by IL-23 and CD11c in skin tissue sections from IMQ treated mice (magnification: 200×); scale bar = 100 μm. Immunostaining with IL-23 antibody was shown as red fluorescence. Immunostaining using antibodies against CD11c was shown as green fluorescence. Nuclei of the cells were localized by DAPI (blue). The orange signals in the merged picture, some of which were shown by red arrows, indicate the co-distribution of IL-23 and CD11c in the cytoplasm of cells. (B) Immunohistochemistry analysis of skin tissue sections using TSLP antibody. Day 0 indicates that the samples were made before the first IMQ application. (C) Daily serum levels of TSLP during the 7-day IMQ treatment as determined by ELISA. (D) Western blot analysis of BRAP expression in HaCaT cells transfected with siRNA to silence C6orf89 expression. (E) ELISA analysis of TSLP content in the culture media of HaCaT cells with C6orf89 expression silenced by siRNA transfection.

Article Snippet: As illustrated by IHC analysis using an antibody against human BRAP (EPR13621, cat: ab181073, Abcam), BRAP was expressed in various cells from both epidermis and dermis.

Techniques: Expressing, Double Immunofluorescence Staining, Immunostaining, Fluorescence, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection

Journal: International Journal of Medical Sciences

Article Title: Bombesin receptor-activated protein homolog deficiency altered the pattern of pathological changes of psoriasis - like skin lesion in mice

doi: 10.7150/ijms.89492

Figure Lengend Snippet: The expression of BRAP and its mouse homolog in skin tissues and the skin injury induced by IMQ in both BC004004 -/- mice and their wild type control mice. (A) The expression of BRAP in both normal human skin and the skin of patient with psoriasis by immunohistochemistry analysis of tissue samples with anti-BRAP antibody. BRAP is expressed in the epidermis of both normal skin tissue and skin tissue with psoriasis (magnification: 100×(scale bar = 200 μm) and 400×(scale bar = 50 μm)). (B) BRAP homologous protein was present in skin tissues as revealed by Western blotting in wild-type control mice BC004004 +/+ with imiquimod (IMQ) treatment for 7 day. The total protein extract from one wild type mice skin representing each day of IMQ treatment was loaded in one well. The expression of β-actin from the same sample was used as a loading control. (C) Representative photographs of psoriasis-like lesions on back skin of BC004004 +/+ mice and BC004004 -/- mice after 7 days of IMQ treatment. Vaseline treatment was applied on control groups as a vehicle control. (D) Daily body weights throughout the 7-day IMQ treatment were shown in the upper left panel. The characteristics of skin lesion were depicted as redness, scaling scores and thickness of back skin and scored according to a murine Psoriasis Area and Severity Index (PASI) score table. Data are presented as mean ± SD; n = 9 in each group. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

Article Snippet: The following antibodies and their corresponding dilutions were used: BRAP (Abcam, cat: ab181073; 1:2,000), IL-23 (Santa Cruz, cat: sc-271279,1:1,000;), IL-1β (Santa Cruz, cat: 12742,1:1,000), IL-10 (Santa Cruz, cat: 8438, 1:1,000), IL-17 (Affinity, cat: DF6127, 1:2,000), IL-17 (Santa Cruz, cat: sc-374218, 1:1,000), TNF-α (Santa Cruz, cat: sc-52746, 1:1,000), CK5 (Affinity, cat: AF5479, 1:2,000), β-Actin (Santa Cruz, cat: sc-47778, 1:5,000), CK14 (Santa Cruz, cat: sc-53253, 1:1,000), TGF-β1 (Abcam, cat: ab64715, 1:2,000), HRP-conjugated goat anti-Mouse IgG (H+L) (Proteintech, cat: SA00001-1, 1:5,000), HRP-conjugated goat anti-rabbit IgG(H+L) (SouthernBiotech, cat: 4050-05; 1: 5,000).

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Bombesin receptor-activated protein homolog deficiency altered the pattern of pathological changes of psoriasis - like skin lesion in mice

doi: 10.7150/ijms.89492

Figure Lengend Snippet: TSLP expression was increased by down-regulation of BRAP or its mouse homolog. (A) Double immunofluorescence staining by IL-23 and CD11c in skin tissue sections from IMQ treated mice (magnification: 200×); scale bar = 100 μm. Immunostaining with IL-23 antibody was shown as red fluorescence. Immunostaining using antibodies against CD11c was shown as green fluorescence. Nuclei of the cells were localized by DAPI (blue). The orange signals in the merged picture, some of which were shown by red arrows, indicate the co-distribution of IL-23 and CD11c in the cytoplasm of cells. (B) Immunohistochemistry analysis of skin tissue sections using TSLP antibody. Day 0 indicates that the samples were made before the first IMQ application. (C) Daily serum levels of TSLP during the 7-day IMQ treatment as determined by ELISA. (D) Western blot analysis of BRAP expression in HaCaT cells transfected with siRNA to silence C6orf89 expression. (E) ELISA analysis of TSLP content in the culture media of HaCaT cells with C6orf89 expression silenced by siRNA transfection.

Article Snippet: The following antibodies and their corresponding dilutions were used: BRAP (Abcam, cat: ab181073; 1:2,000), IL-23 (Santa Cruz, cat: sc-271279,1:1,000;), IL-1β (Santa Cruz, cat: 12742,1:1,000), IL-10 (Santa Cruz, cat: 8438, 1:1,000), IL-17 (Affinity, cat: DF6127, 1:2,000), IL-17 (Santa Cruz, cat: sc-374218, 1:1,000), TNF-α (Santa Cruz, cat: sc-52746, 1:1,000), CK5 (Affinity, cat: AF5479, 1:2,000), β-Actin (Santa Cruz, cat: sc-47778, 1:5,000), CK14 (Santa Cruz, cat: sc-53253, 1:1,000), TGF-β1 (Abcam, cat: ab64715, 1:2,000), HRP-conjugated goat anti-Mouse IgG (H+L) (Proteintech, cat: SA00001-1, 1:5,000), HRP-conjugated goat anti-rabbit IgG(H+L) (SouthernBiotech, cat: 4050-05; 1: 5,000).

Techniques: Expressing, Double Immunofluorescence Staining, Immunostaining, Fluorescence, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection

Journal: Stress (Amsterdam, Netherlands)

Article Title: Lack of bombesin receptor-activated protein homologous protein impairs hippocampal synaptic plasticity and promotes chronic unpredictable mild stress induced behavioral changes in mice.

doi: 10.1080/10253890.2022.2155513

Figure Lengend Snippet: Figure 2. The analysis of behavioral tests after exposure to chronic unpredictable mild stress (CUMS). (A) Body weight changes throughout the CUMS procedure. (B) Immobility time of bc004004þ/þ and bc004004 / mice in the forced swimming test with treatment of CUMS. (C) Analysis of sucrose preference tests during the CUMS procedure. (D) Immobility time of bc004004þ/þ and bc004004 / mice in the tail suspension test after 21-day CUMS exposure. (E) Total distance traveled in the different regions in the open field test after 21-day CUMS exposure. Two-way ANOVA was performed followed by Tukey’s multiple comparisons. (F) The expres sion of BRAP homologous protein in mice detected by western blot with anti-BRAP antibody. The protein extracts of hippocampus tissues of the mice was used in western blot after 21-day of CUMS treatment. The expression of b-actin from the same sample was used as loading controls. Data are presented as mean ± SD. p <.05, p <.01, p <.001,p <.0001, n ¼ 8.

Article Snippet: The transferred PVDF membranes were incubated with the following primary antibodies with dilutions indicated respectively: b-actin (Sigma, cat: A5441, 1:10,000), BDNF (Abcam, cat: ab108319, 1:1000), Trk-B (Boster, cat: BA0623, 1:1000), BRAP (Abcam, cat: ab181073, 1:1000), GluN2A (CUSABIO, cat: CSB-PA14129A0Rb, 1:500), PSD95 (Sangon Biotech, cat: D261092, 1:500), synaptophysin (ABclonal, cat: A6344, 1:1000).

Techniques: Suspension, Western Blot, Expressing