Journal: Scientific Reports

Article Title: Janus-faced Acrolein prevents allergy but accelerates tumor growth by promoting immunoregulatory Foxp3+ cells: Mouse model for passive respiratory exposure

doi: 10.1038/srep45067

Figure Lengend Snippet: ( A ) Mice nasally treated with KLH (K), acrolein ( A ), PBS (P) or with KLH in combination with acrolein (KA) were exposed to increasing doses of methacholine and the degree of airway hyperreactivity was measured by whole body plethysmography according to change in area of enhanced pause (Penh). ( B ) Rectal temperature drop 20 min after ip-challenge with KLH as a parameter to determine systemic anaphylactic reactions ( C ) Summary of CD4+CD25+Foxp3+ cell numbers counted in the spleens of the different treated mice of two independently performed experiments. ( D) Whole lung sections were imaged by TissueFAX and positive stainings were quantified by Histoquest. Summary of immune-histochemical quantification % of CD3+ cells, ( D ) % of Foxp3+ cells ( E ) Foxp3/CD3 ratio of lung sections in the different treated groups. ( F ) Representative regions of lung sections stained for CD3 and Foxp3. 2 stained sections per animal were analysed by TissueFAX. Groups were compared by ANOVA following Newman-Keuls Multiple Comparison test. Mean ± SEM; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: After cooling down slides were blocked for endogenous peroxidase (3% H 2 O 2 in Methanol, 10 min), permeabilization with 0.2% Tween in PBS and for unspecific binding with 1.5% blocking serum (rabbit ABC staining system, Santa Cruz) and incubated with rabbit anti-mouse Foxp3 (1:400 in 1.5% goat serum)(1.98 mg/ml) or isotype control, rabbit IgG, overnight at 4 °C.

Techniques: Staining, Comparison

Journal: Scientific Reports

Article Title: Janus-faced Acrolein prevents allergy but accelerates tumor growth by promoting immunoregulatory Foxp3+ cells: Mouse model for passive respiratory exposure

doi: 10.1038/srep45067

Figure Lengend Snippet: ( A ) As depicted in the treatment protocol, mice were intranasally pretreated 6 times in biweekly intervals with acrolein or PBS. After one week interval, murine mammary carcinoma cell-line D2F2 derived from BALB/c mice were injected subcutaneously in the right flank and their growth was monitored. ( B ) Representative pictures of excised tumors of mice previously pre-treated intranasally with PBS (P, n = 8), or acrolein (A, n = 8). Summary of ( C ) tumor size and ( D ) body weight of mice 11 days post-tumor engraftment in the different groups. Whole tumor sections were imaged by TissueFAX and positive stainings were quantified by Histoquest. Summary of immune-histochemical quantification of ( E ) CD3+ cells, ( F ) Foxp3+ cells and ( G ) Foxp3/CD3 ratio of tumor sections in the different treated groups. ( H ) Representative regions of tumor sections stained for CD3, Foxp3 and their respective isotype controls are shown. 2 sections/animal and staining were analyzed by Tissue FAX. Groups were compared by unpaired T test. Mean ± SEM; * P < 0.05; ** P < 0.01.

Article Snippet: After cooling down slides were blocked for endogenous peroxidase (3% H 2 O 2 in Methanol, 10 min), permeabilization with 0.2% Tween in PBS and for unspecific binding with 1.5% blocking serum (rabbit ABC staining system, Santa Cruz) and incubated with rabbit anti-mouse Foxp3 (1:400 in 1.5% goat serum)(1.98 mg/ml) or isotype control, rabbit IgG, overnight at 4 °C.

Techniques: Derivative Assay, Injection, Staining

Journal: Scientific Reports

Article Title: Janus-faced Acrolein prevents allergy but accelerates tumor growth by promoting immunoregulatory Foxp3+ cells: Mouse model for passive respiratory exposure

doi: 10.1038/srep45067

Figure Lengend Snippet: ( A ) NF-κB inhibition was measured with monocytic THP1-XBlue cells, which were treated with LPS and increasing concentrations of acrolein or cinnamaldehyde for 18 hours. Bars represent summary from two independent experiments normalized to cells stimulated with LPS alone. ( B ) AZ-AhR cells were treated with different concentrations of acrolein or cinnamaldehyde crotonaldehyde, propanal and methacrolein in increasing concentrations (0.018/0.18/1.8/9/18/45/90/180 μM), with ( C ) acrolein alone or in combination with the antagonists resveratrol or 3′-methoxy-4′-nitroflavone for 18 h, before luciferase-activity was measured in the supernatant. Bars represent data from three independent experiments normalized to medium alone for B and two independent experiments for C. Data presented as mean ± SD, Statistical analyses was made using 2way RM ANOVA following Sidak’s multiple comparisons test, respectively. Significances to controls are depicted. ( D ) PBMCs were stimulated for 72 h with cinnamaldehyde, acrolein and or resveratrol. Gating Strategy and summary of % of CD4+CD25+Foxp3+ cells as determined by FACS of 5 independent experiments (n = 15). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001.

Article Snippet: After cooling down slides were blocked for endogenous peroxidase (3% H 2 O 2 in Methanol, 10 min), permeabilization with 0.2% Tween in PBS and for unspecific binding with 1.5% blocking serum (rabbit ABC staining system, Santa Cruz) and incubated with rabbit anti-mouse Foxp3 (1:400 in 1.5% goat serum)(1.98 mg/ml) or isotype control, rabbit IgG, overnight at 4 °C.

Techniques: Inhibition, Luciferase, Activity Assay