Journal: International Journal of Molecular Medicine

Article Title: The interplay of BMP4 and IL-7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia/reperfusion

doi: 10.3892/ijmm.2018.3480

Figure Lengend Snippet: The bone morphogenetic protein (BMP) signaling pathway is activated in intestinal epithelial cells and intraepithelial lymphocytes (IELs) following ischemia/reperfusion (I/R). (A) The level of BMP4 protein (red) expression significantly increased in the mid-to-distal villus region after 6 h of I/R, according to immunofluorescence staining. (B) The expression levels of the type I BMP receptor and phosphorylated nuclear factor (p-NF)-κB were both significantly increased after 6 h of I/R in IELs. * P<0.05, 3-4 mice/group; the results are representative of three independent experiments.

Article Snippet: Recombinant BMP4 protein (cat. no. 315-27; 30 ng/ml) and recombinant murine IL-7 protein (cat. no. 217-17; 2 ng/ml) were both purchased from ProteinTech Group, Inc. Pyrrolidine dithiocarbamate (PDTC; cat. no. S1808; 30 ng/ml), an inhibitor of NF-κB, was purchased from Beyotime Institute of Biotechnology (Wuhan, China).

Techniques: Expressing, Immunofluorescence, Staining

Journal: International Journal of Molecular Medicine

Article Title: The interplay of BMP4 and IL-7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia/reperfusion

doi: 10.3892/ijmm.2018.3480

Figure Lengend Snippet: Activation of nuclear factor (NF)-κB signaling by bone morphogenetic protein (BMP)4 stimulation in intraepithelial lymphocytes (IELs). Flow cytometry determined the expression of the BMP type I receptor and phosphorylated NF-κB in IELs in culture. Following treatment with BMP4 for 6 h, the expression of the BMP type I receptor and phosphorylated NF-κB was significantly increased compared with that in the control group. NOGGIN partially decreased the expression of the BMP type I receptor, as well as NF-κB transcriptional activity. The results are representative of three independent experiments. P<0.05.

Article Snippet: Recombinant BMP4 protein (cat. no. 315-27; 30 ng/ml) and recombinant murine IL-7 protein (cat. no. 217-17; 2 ng/ml) were both purchased from ProteinTech Group, Inc. Pyrrolidine dithiocarbamate (PDTC; cat. no. S1808; 30 ng/ml), an inhibitor of NF-κB, was purchased from Beyotime Institute of Biotechnology (Wuhan, China).

Techniques: Activation Assay, Flow Cytometry, Expressing, Control, Activity Assay

Journal: International Journal of Molecular Medicine

Article Title: The interplay of BMP4 and IL-7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia/reperfusion

doi: 10.3892/ijmm.2018.3480

Figure Lengend Snippet: Bone morphogenetic protein (BMP)4 induces intraepithelial lymphocytes (IELs) to undergo apoptosis. Intestinal IELs were examined by flow cytometry for markers of apoptosis (FITC-Annexin V and PI). FITC-Annexin V + /PI + indicates late apoptosis, FITC-Annexin V + /PI − indicates early apoptosis, and FITC-Annexin V − /PI − indicates live cells. The extent of apoptosis of IELs after treatment with BMP4 for 6 h was then determined. The expression of FITC-Annexin V + /PI + IELs in the BMP4 group was significantly higher compared with that in the control group, but these effects were decreased by treatment with NOGGIN or pyrrolidine dithiocarbamate (PDTC). P<0.05, 3-4 mice/group; each experiment was repeated three times.

Article Snippet: Recombinant BMP4 protein (cat. no. 315-27; 30 ng/ml) and recombinant murine IL-7 protein (cat. no. 217-17; 2 ng/ml) were both purchased from ProteinTech Group, Inc. Pyrrolidine dithiocarbamate (PDTC; cat. no. S1808; 30 ng/ml), an inhibitor of NF-κB, was purchased from Beyotime Institute of Biotechnology (Wuhan, China).

Techniques: Flow Cytometry, Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: The interplay of BMP4 and IL-7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia/reperfusion

doi: 10.3892/ijmm.2018.3480

Figure Lengend Snippet: Inhibition of bone morphogenetic protein (BMP)4 induces the apoptosis of intraepithelial lymphocytes (IELs) that has been stimulated by interleukin (IL)-7. Flow cytometry and apoptosis markers (FITC-Annexin V and PI) were used to examine IEL apoptosis after treatment with BMP4, IL-7 or BMP4 + IL-7 for 6 h. The expression of FITC-Annexin V + /PI + IELs in the IL-7 treatment group was significantly lower compared with that in the control group, whereas the expression of FITC-Annexin V + /PI + IELs in the BMP4 treatment group was significantly higher compared with that in the control group. However, the expression of FITC-Annexin V + /PI + IELs in the BMP4 + IL-7 treatment group exhibited no changes compared with the control group. Each experiment was repeated three times; 4-5 mice/group. P<0.05.

Article Snippet: Recombinant BMP4 protein (cat. no. 315-27; 30 ng/ml) and recombinant murine IL-7 protein (cat. no. 217-17; 2 ng/ml) were both purchased from ProteinTech Group, Inc. Pyrrolidine dithiocarbamate (PDTC; cat. no. S1808; 30 ng/ml), an inhibitor of NF-κB, was purchased from Beyotime Institute of Biotechnology (Wuhan, China).

Techniques: Inhibition, Flow Cytometry, Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: The interplay of BMP4 and IL-7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia/reperfusion

doi: 10.3892/ijmm.2018.3480

Figure Lengend Snippet: Interleukin (IL)-7 downregulates the bone morphogenetic protein (BMP) signaling pathway in intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs). (A) Western blot analysis was used to determine the expression of BMP4 in IEC-6 cells following treatment with IL-7 for 6 h. The expression of BMP4 was significantly decreased compared with that in the control group. (B) Flow cytometry, was used to detect the phosphorylation of nuclear factor (NF)-κB following treatment with IL-7 for 6 h. The expression of phosphorylated NF-κB was significantly decreased compared with that in the control group. Each experiment was repeated three times; 4-5 mice/group. * P<0.05 vs. control. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Recombinant BMP4 protein (cat. no. 315-27; 30 ng/ml) and recombinant murine IL-7 protein (cat. no. 217-17; 2 ng/ml) were both purchased from ProteinTech Group, Inc. Pyrrolidine dithiocarbamate (PDTC; cat. no. S1808; 30 ng/ml), an inhibitor of NF-κB, was purchased from Beyotime Institute of Biotechnology (Wuhan, China).

Techniques: Western Blot, Expressing, Control, Flow Cytometry, Phospho-proteomics

Journal: International Journal of Molecular Medicine

Article Title: The interplay of BMP4 and IL-7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia/reperfusion

doi: 10.3892/ijmm.2018.3480

Figure Lengend Snippet: Bone morphogenetic protein (BMP)4 regulates the interleukin (IL)-7α/CD127 signaling pathway in intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs). (A) Western blot analysis determined the expression of IL-7 in IEC-6 cells following treatment with BMP4 for 6 h. The IL-7 level was significantly decreased compared with that in the control group. (B) Flow cytometry, was used to detect the expression of CD127 and phosphorylated signal transducer and activator of transcription (STAT)5 proteins following treatment with BMP4 for 6 h. The levels of CD127 and phosphorylated STAT5 proteins were significantly decreased compared with those in the control group. Each experiment was repeated three times; 4-5 mice/group. * P<0.05 for control vs. treatment with BMP4 alone; ** P<0.05 for control vs. the BMP4 + NOGGIN group. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Recombinant BMP4 protein (cat. no. 315-27; 30 ng/ml) and recombinant murine IL-7 protein (cat. no. 217-17; 2 ng/ml) were both purchased from ProteinTech Group, Inc. Pyrrolidine dithiocarbamate (PDTC; cat. no. S1808; 30 ng/ml), an inhibitor of NF-κB, was purchased from Beyotime Institute of Biotechnology (Wuhan, China).

Techniques: Western Blot, Expressing, Control, Flow Cytometry

Journal: STAR Protocols

Article Title: Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells

doi: 10.1016/j.xpro.2022.101785

Figure Lengend Snippet:

Article Snippet: Download the appropriate sequences. a. Download the Reference Sequence for Homo sapiens bromodomain containing 4 (BRD4), transcript variant long, mRNA (GenBank: NM_058243.3) from the NCBI’s website, using the GenBank data base ( BRD4-L RefSeq ). b. Download the sequence for LentiV_Blast (Addgene, cat# 111887) from Addgene’s website ( LentiV_Blast ).

Techniques: Virus, Recombinant, Cloning, Gel Extraction, Plasmid Preparation, Sequencing, Over Expression, Expressing, Software, Imaging