th9 induction medium  (Proteintech)

Journal: International immunopharmacology

Article Title: Foxp2 inhibits Th9 cell differentiation and attenuates allergic airway inflammation in a mouse model of ovalbumin-induced asthma.

doi: 10.1016/j.intimp.2022.109060

Figure Lengend Snippet: Fig. 1. Anti-IL-9 antibody inhibits Th9 cells differentiation in asthmatic mice. Forty BALB/c mice were randomly divided into four groups: control group (Control), OVA-induced asthma model group (OVA), mouse IgG injection treatment group (IgG) and anti-IL-9 mAb injection treatment group (anti-IL-9). Four hours after the last OVA stimulation, the mice were anesthetized and killed to collect bronchoalveolar lavage fluid (BALF) and lung tissue. A-B. The percentage of Th9 cells in BALF was evaluated by flow cytometry in each group. C-D. The percentage of Th9 cells in lung tissue was evaluated by flow cytometry in each group. E-F. The levels of IL-9 in BALF and lung tissue were measured by ELISA in each group. G-H. The mRNA expression of BATF and IRF4 in lung tissue were detected by RT-PCR. I-K. The protein expression of BATF and IRF4 in lung tissue were detected by western blot. **P < 0.01,***p < 0.001 vs Control group, #P < 0.05, ##P < 0.01,###p < 0.001 vs IgG group.

Article Snippet: Subsequently, CD4+ T cells were cultured in Th9 induction medium (10 ng/mL IL-4, 3 ng/mL TGF-β, and 10 mg/mL anti-IFN-γ, Proteintech).

Techniques: Control, Injection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: International immunopharmacology

Article Title: Foxp2 inhibits Th9 cell differentiation and attenuates allergic airway inflammation in a mouse model of ovalbumin-induced asthma.

doi: 10.1016/j.intimp.2022.109060

Figure Lengend Snippet: Fig. 3. Down regulation of Foxp2 expression in Th9 cells Spleen cells were isolated from normal mice and CD4+T cells were sorted by magnetic beads. IL-4, anti–IFN-ɣ and TGF-β were used to induce the differentiation of Th9 cells. After 4 days of induction, cells were collected. A.The levels of IL-9 in the supernatant of pre-induction (CD4+T) and post-induction (Th9) cells were were measured by ELISA. B-C. The mRNA expres sion of IL-9 and Foxp2 in pre-induction (CD4+T) and post-induction (Th9) cells were detected by RT-PCR in each group. D-E. The protein expression of IL-9 and Foxp2 in pre-induction (CD4+T) and post-induction (Th9) cells were analyzed by western blot in each group. **P < 0.01,***p < 0.001 vs CD4 + T cell group.

Article Snippet: Subsequently, CD4+ T cells were cultured in Th9 induction medium (10 ng/mL IL-4, 3 ng/mL TGF-β, and 10 mg/mL anti-IFN-γ, Proteintech).

Techniques: Expressing, Isolation, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: International immunopharmacology

Article Title: Foxp2 inhibits Th9 cell differentiation and attenuates allergic airway inflammation in a mouse model of ovalbumin-induced asthma.

doi: 10.1016/j.intimp.2022.109060

Figure Lengend Snippet: Fig. 4. Overexpression of Foxp2 inhibits Th9 cell differentiation in vitro Th9 cells were infected withFoxp2 overexpressed lentivirus (LV-Foxp2) and control lentivirus (LV-NC), respectively, and no infected group as control. A-B. The percentage of Th9 cells was evaluated by Flow cytometry. C. The levels of IL-9 in cell supernatant was measured by ELISA. D. The mRNA expression of IL-9 in cells was detected by RT-PCR. E-F. The mRNA expression of BATF and IRF4 in cells were detected by RT- PCR. **P < 0.01, ***p < 0.001 vs LV-NC group.

Article Snippet: Subsequently, CD4+ T cells were cultured in Th9 induction medium (10 ng/mL IL-4, 3 ng/mL TGF-β, and 10 mg/mL anti-IFN-γ, Proteintech).

Techniques: Over Expression, Cell Differentiation, In Vitro, Infection, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: International immunopharmacology

Article Title: Foxp2 inhibits Th9 cell differentiation and attenuates allergic airway inflammation in a mouse model of ovalbumin-induced asthma.

doi: 10.1016/j.intimp.2022.109060

Figure Lengend Snippet: Fig. 5. Overexpression of Foxp2 improves airway inflammation in asthmatic mice by inhibiting Th9 cell differentiation. Ten BALB/c mice were fed adaptively for one week. In the second week, 10 mice were divided into two groups (OVA + NC group and OVA + Foxp2 group) to establish asthma model. In OVA + NC group, OVA sensitized/challenged asthmatic mice were given control lentivirus by tracheal delivery at 3 days before aerosol challenge. In OVA + Foxp2 group, OVA sensitized/challenged asthmatic mice were given Foxp2 overexpression lentivirus by tracheal delivery at 3 days before aerosol challenge. Four hours after the last challenge, the mice were anesthetized and killed to get the lung tissue. A. HE staining was used to observe the pathological changes of lung tissue. B. PAS glycogen staining was used to determine the airway mucus secretion. C-F. The mRNA expression of IL-9, FOXP2, BATF and IRF4 in lung tissue were detected by RT-PCR. *P < 0.05,**P < 0.01,***p < 0.001 vs OVA + NC group.

Article Snippet: Subsequently, CD4+ T cells were cultured in Th9 induction medium (10 ng/mL IL-4, 3 ng/mL TGF-β, and 10 mg/mL anti-IFN-γ, Proteintech).

Techniques: Over Expression, Cell Differentiation, Control, Aerosol, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction