Journal: Respiratory research

Article Title: Combined genomic and proteomic approaches reveal DNA binding sites and interaction partners of TBX2 in the developing lung.

doi: 10.1186/s12931-021-01679-y

Figure Lengend Snippet: Fig. 5 Scheme of the molecular interactions of TBX2 in repression of target genes in the pulmonary mesenchyme. Grey discs represent nucleosomes. DNA is displayed as black line. Dotted line indicates unknown length of DNA fragment and possible long distance. Circles for proteins do not represent the real size. TBX2 binds in concert with HMGB2 and PBX1 to specific DNA sites in enhancer regions. Binding to PBX1 is probably indirect via unknown proteins, binding to HMGB2 is direct and may aid in chromatin bending and bringing enhancer regions to the proximity of transcription start sites. TBX2/DNMT1 binding may induce DNA methylation to repress gene expression. TBX2 and CBX3 directly bind and both associate with the NuRD complex for chromatin remodeling. TBX2/MYBBP1 interaction may strengthen the association with the NuRD complex to repress target genes. TBX2 directly interacts with components of the NuRD complex, namely HDACs for chromatin deacetylation and the chromatin remodeling complex CHD4. TBX2 interaction with CBX3, HDACs and CHD4 collectively induces chromatin compaction. RBBP4/7 binds to chromatin to consolidate binding of the NuRD-complex to target gene DNA

Article Snippet: Pbx1b encoding full-length mouse PBX1B, (gift from Heike Pöpperl, Institute for Biophysical Chemistry, Hannover Medical School, Germany); pd2EGFP-N1 (EGFPexpression vector) [35]; pCMV6-Entry.Cbx3-Myc-DDK encoding full-length mouse CBX3 (#MR224357, OriGene, Rockville, Maryland, USA); pCMV-SPORT6.Hdac1 encoding full-length mouse HDAC1 (#4217199, Sourcebioscience, Nottingham, UK); pCMV6-Entry.Hdac2-MycDDK encoding full-length mouse HDAC2 (#MR226709, OriGene); pCMV-SPORT6.Chd4 encoding full-length mouse CHD4 (#6489649, Sourcebioscience); pCMVEntry.Hmgb2-GFP encoding full-length mouse HMGB2 (#MR202276, OriGene).

Techniques: Binding Assay, DNA Methylation Assay, Gene Expression

Journal: Scientific Reports

Article Title: Evidence for a non-stochastic two-field hypothesis for persistent skin cancer risk

doi: 10.1038/s41598-020-75864-2

Figure Lengend Snippet: Persistent hyperemic foci exhibit evidence of dermal senescence. ( A – E ) Dermal cells in areas of high Hgb content exhibit a heterochromatin pattern of DAPI staining. After excision of skin from mice treated with and without UV at 2 and 20 weeks after stopping UV treatments, the sections were stained with DAPI to better demonstrate nuclear morphology. Nuclei of dermal cells showing a heterochromatin staining pattern are shown by white arrows while nuclei exhibiting a euchromatin pattern of DAPI staining are shown by yellow arrows. The hatched white line outlines the epidermal-dermal junction. The white scale bar represents 50 µm. ( A ) Skin from an area of low Hgb content 2 weeks after stopping UV treatments. ( B ) Skin from an area of high Hgb content 2 weeks after stopping UV treatments. ( C ) Control non-UV treated skin. ( D ) Skin from an area of low Hgb content 20 weeks after stopping UV. ( E ) Skin from an area of high Hgb content 20 weeks after stopping UV. (F – J ) Representative photomigrographs of skin following immunolabeling with anti-p16 INK4a antibodies. ( F ) Control non-UV treated epidermis. ( G , H ) Skin excised 2 weeks after stopping UV treatments from an area of low Hgb content ( G ) or from an area of high Hgb content ( H ). ( I , J ) Skin excised from a low Hgb area ( I ) or a high Hgb area ( J ) at 20 weeks after stopping UV treatments. Black arrows in ( H , J ) show dermal cells with enlarged nuclei labeling positive for p16 INK4a . The black scale bars represent 100 µm. ( K ) HP1γ + dermal cells are increased in hyperemic areas at both 2 and 20 weeks after stopping UV treatments. Immunofluorescent (IF) labeling of formalin-fixed skin sections was performed using anti-HP1γ and anti-pancytokeratin (CK) antibody. The data depicts the % of dermal cells positive for HP1γ nuclear labeling. ( L ) Dermal cells positive for nuclear γH2AX are also increased in hyperemic areas only at 2 and 20 weeks post-UV. IF was performed for both nuclear γH2AX immunolabeling and CK. The data shown is the percentage of γH2AX + dermal cells relative to all CK negative cells. (ns = non-significant; ** = p < 0.01; *** = p < 0.001).

Article Snippet: Immunofluorescent staining of formalin-fixed paraffin-embedded mouse skin following heat-induced antigen retrieval was performed using the following antibodies: HP1γ (Cbx3) immunolabeling was done using rabbit anti-Cbx3 (1:250, Cat#IHC-00204, Bethyl Laboratories). γH2AX immunolabeling was done using monoclonal rabbit anti-γH2AX (1:100, Bethyl Laboratories, Cat#A700-053).

Techniques: Staining, Control, Immunolabeling, Labeling