Journal: Developmental Cell

Article Title: Defining the identity and the niches of epithelial stem cells with highly pleiotropic multilineage potency in the human thymus

doi: 10.1016/j.devcel.2023.08.017

Figure Lengend Snippet:

Article Snippet: HumanKine® recombinant human FLT3 Ligand protein , Proteintech , cat# HZ-1151.

Techniques: Recombinant, Modification, Selection, RNA Extraction, Polymer, Cell Culture, Derivative Assay, Software

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Regulation of FLT3 signaling in lung tissue by FLT3L and lestaurtinib. mRNA and protein expression levels of Akt, ERK1/2 and STAT5 were measured by reverse transcription-quantitative PCR and western blotting, respectively, to confirm regulation of FLT3 signaling by FLT3L and lestaurtinib. (A) mRNA expression of Akt. (B) mRNA expression of ERK1/2. (C) mRNA expression of STAT5. β-actin was used as a reference gene. (D) Western blot analysis of Akt, ERK1/2 and STAT5 and their phosphorylated forms. (E) Ratio of p-Akt to total Akt levels. (F) Ratio of p-ERK to total ERK levels. (G) Ratio of p-STAT5 to total STAT5 levels. β-actin was used as a reference protein. Data are presented as the mean ± standard deviation (n=3). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; and $ P<0.05 vs. lestaurtinib+ARDS. FLT3, Fms-like tyrosine kinase 3; FLT3L, Fms-like tyrosine kinase 3-ligand; p-, phosphorylated; ARDS, acute respiratory distress syndrome.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Effect of FLT3 signaling on the aggregation and maturation of pulmonary cDCs. The aggregation and maturation of pulmonary cDCs were measured by flow cytometry at 6 h post-LPS challenge. (A) Representative dot plots and (B) quantification showing the percentage of cDCs (CD11c + CD11b + ) among total lung cells in the experimental groups. (C) Representative histograms and (D) quantification showing the expression of MHC II in pulmonary cDCs in the experimental groups. (E) Representative histograms and (F) quantification showing the expression of CD80 in pulmonary cDCs in the experimental groups. Data are presented as the mean ± standard deviation (n=6). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; and $ P<0.05 vs. lestaurtinib+ARDS. FLT3, Fms-like tyrosine kinase 3; cDCs, classical dendritic cells; LPS, lipopolysaccharide; MHC II, major histocompatibility complex class II; ARDS, acute respiratory distress syndrome; FLT3L, Fms-like tyrosine kinase 3-ligand.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Flow Cytometry, Expressing, Standard Deviation, Control, Immunopeptidomics

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Effect of cDC manipulation on acute lung inflammation in ARDS. (A) Levels of TNF-α and (B) IL-6 were measured using ELISA to evaluate acute lung inflammation at 6 h post-LPS challenge. Data are presented as the mean ± standard deviation (n=6). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; and $ P<0.05 vs. lestaurtinib+ARDS. cDC, classical dendritic cell; ARDS, acute respiratory distress syndrome; TNF, tumor necrosis factor; IL, interleukin; LPS, lipopolysaccharide; FLT3L, Fms-like tyrosine kinase 3-ligand.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Effect of cDC manipulation on lung edema and lung injury. LWW/BW and the lung injury score were measured to assess lung edema and lung injury at 6 and 24 h post-LPS challenge, respectively. (A) Hematoxylin and eosin staining (magnification, ×200) of lung tissues at 6 h and (B) at 24 h post-LPS challenge. (C) Lung injury score at 6 h and (D) at 24 h post-LPS challenge. (E) LWW/BW at 6 h and (F) at 24 h post-LPS challenge. Lung injury score is expressed as arbitrary mean units. Data are presented as the mean ± standard deviation (n=6). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; and $ P<0.05 vs. lestaurtinib+ARDS. cDC, classical dendritic cell; LWW/BW, lung wet weight to body weight; LPS, lipopolysaccharide; ARDS, acute respiratory distress syndrome; FLT3L, Fms-like tyrosine kinase 3-ligand.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Staining, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Effect of cDC manipulation on neutrophil infiltration. MPO activity was measured by chromometry to assess neutrophil infiltration (A) at 6 h and (B) at 24 h post-LPS challenge. MPO activity is expressed as units per gram of sample. Data are presented as the mean ± standard deviation (n=6). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; $ P<0.05 vs. lestaurtinib+ARDS. cDC, classical dendritic cell; MPO, myeloperoxidase; LPS, lipopolysaccharide; ARDS, acute respiratory distress syndrome; FLT3L, Fms-like tyrosine kinase 3-ligand.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Activity Assay, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Effect of cDC manipulation on the balance of the Th1/Th2 response. The mRNA and protein expression levels of T-bet and GATA-3 were measured by reverse transcription-quantitative PCR and western blotting to evaluate the balance of the Th1/Th2 response. (A) mRNA expression of T-bet and (B) GATA-3. β-actin was used as a reference gene. (C) Representative blots from western blotting analysis of T-bet and GATA-3 protein expression. (D) Protein expression levels of T-bet and (E) GATA-3. β-actin was used as a reference protein. Data are presented as the mean ± standard deviation (n=3). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; and $ P<0.05 vs. lestaurtinib+ARDS. cDC, classical dendritic cell; Th, T helper cell; T-bet, T-box-expressed-in-T-cells; GATA-3, GATA binding protein 3; ARDS, acute respiratory distress syndrome; FLT3L, Fms-like tyrosine kinase 3-ligand.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, Control, Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: Effect of cDC manipulation on Th1/Th2 cytokine production. (A) Levels of IFN-γ, (B) IL-1β, (C) IL-4 and (D) IL-10 were measured in the lungs by ELISA at 24 h post-LPS challenge, in order to evaluate the Th1/Th2 cytokine production. Data are presented as the mean ± standard deviation (n=6). * P<0.05 vs. Control; # P<0.05 vs. ARDS; & P<0.05 vs. FLT3L+ARDS; and $ P<0.05 vs. lestaurtinib+ARDS. cDC, classical dendritic cell; Th, T helper cell; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; ARDS, acute respiratory distress syndrome; FLT3L, Fms-like tyrosine kinase 3-ligand.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Classical dendritic cells regulate acute lung inflammation and injury in mice with lipopolysaccharide-induced acute respiratory distress syndrome

doi: 10.3892/ijmm.2019.4208

Figure Lengend Snippet: FLT3L expression in normal and ARDS lung tissues. FLT3L protein expression levels in normal and ARDS lung tissues were evaluated by western blotting at 6 h post-LPS challenge. Data are presented as the mean ± standard deviation (n=3). * P<0.05 vs. Control. FLT3L, Fms-like tyrosine kinase 3-ligand; ARDS, acute respiratory distress syndrome.

Article Snippet: After SDS-PAGE (10%), the proteins were transferred to a polyvinylidene fluoride membrane, blocked with Tris-buffered saline with Tween-20 (TBST) containing 5% skim milk powder at 25°C for 1 h, and incubated at 4°C overnight with primary antibodies against phosphorylated (p-) Akt, total Akt, p-ERK1/2, total ERK1/2, p-STAT5, total STAT5 (all at 1:400 dilution; cat. nos. ab38449, ab8805, ab201015, ab17942, ab32364, ab16276; Abcam), T-bet, GATA-3, FLT3L and β-actin (all at 1:400 dilution; cat. nos. sc-21763, sc-130057, sc-365266, sc-47778; Santa Cruz Biotechnology, Inc.). β-actin was used as an internal reference protein.

Techniques: Expressing, Western Blot, Standard Deviation, Control