Journal: Cell reports

Article Title: c-FOS drives reversible basal to squamous cell carcinoma transition

doi: 10.1016/j.celrep.2021.109774

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CFOS-FAM: Mm00487425 (TaqMan Gene Expression Assays) , ThermoFisher , N/A.

Techniques: Virus, Plasmid Preparation, Membrane, Recombinant, Purification, Gene Expression, shRNA, Control, Software

Journal: Life

Article Title: Identification of New, Functionally Relevant Mutations in the Coding Regions of the Human Fos and Jun Proto-Oncogenes in Rheumatoid Arthritis Synovial Tissue

doi: 10.3390/life11010005

Figure Lengend Snippet: Cfos and cjun mutations detected in synovial tissue of rheumatoid arthritis (RA) and osteoarthritis (OA) patients.

Article Snippet: For cFos and cJun protein detection, SDS-PAGE (10%) was performed using 10 μL of each whole cell extract sample, followed by Western Blot analysis using the respective cFos and cJun antibodies from the TransAM AP-1 Family Kit (Active Motif, Rixensart, Belgium).

Techniques:

Journal: Life

Article Title: Identification of New, Functionally Relevant Mutations in the Coding Regions of the Human Fos and Jun Proto-Oncogenes in Rheumatoid Arthritis Synovial Tissue

doi: 10.3390/life11010005

Figure Lengend Snippet: Effects of cjun and cfos mutations on reporter gene expression. The graph shows pentameric AP-1 site- and matrix metalloproteinase 1 (MMP-1) promoter-dependent expression of firefly luciferase 2 days following transfection of NIH-3T3 cells with wt ( A ) or mutated ( B , C ) cfos and cjun expression plasmids (determined in biological triplicates; means ± standard error of the mean). Firefly luciferase expression levels were normalized to renilla luciferase expression levels in the respective samples (transfection and normalization control). Results are presented as relative luciferase activity (in x-fold) related to the expression level in the presence of the control vector ( A ), cfos wt expression vector ( B ), or cfos and cjun wt vectors ( C ). For normalization and to correct for different transfection efficiencies, the renilla luciferase-expressing vector pRL-CMV was co-transfected. In all analyzed samples, luciferase expression was easy to detect and clearly exceeded background levels in non-transfected cells. * p ≤ 0.05 vs. controls, i.e., pCMX ( A ), fosWt ( B ), and fosWt/junWt ( C ); +, °, §, ‡, and # p ≤ 0.05 vs. respective other mutants; fos73 represents the double mutant fos73/fos125.

Article Snippet: For cFos and cJun protein detection, SDS-PAGE (10%) was performed using 10 μL of each whole cell extract sample, followed by Western Blot analysis using the respective cFos and cJun antibodies from the TransAM AP-1 Family Kit (Active Motif, Rixensart, Belgium).

Techniques: Gene Expression, Expressing, Luciferase, Transfection, Control, Activity Assay, Plasmid Preparation, Mutagenesis

Journal: Life

Article Title: Identification of New, Functionally Relevant Mutations in the Coding Regions of the Human Fos and Jun Proto-Oncogenes in Rheumatoid Arthritis Synovial Tissue

doi: 10.3390/life11010005

Figure Lengend Snippet: DNA binding activity of AP-1 complexes in transfected K4IM fibroblasts and protein expression of cfos and cjun variants. ( A ) Electrophoretic mobility shift assay (EMSA) of nuclear extracts from transfected K4IM cells. The phosphor-image exemplarily shows binding of radio-labelled AP-1 sites by AP-1 complexes formed in K4IM cells transfected with the indicated cfos and/or cjun variants ( n = 1). ( B ) In the corresponding whole cell extracts, protein amounts of cFos and cJun following transfection were detected by Western Blot ( n = 1). Loading control: glyceraldehyde 3-phosphate dehydrogenase (GAPDH); fos73: double mutant fos73/fos125.

Article Snippet: For cFos and cJun protein detection, SDS-PAGE (10%) was performed using 10 μL of each whole cell extract sample, followed by Western Blot analysis using the respective cFos and cJun antibodies from the TransAM AP-1 Family Kit (Active Motif, Rixensart, Belgium).

Techniques: Binding Assay, Activity Assay, Transfection, Expressing, Electrophoretic Mobility Shift Assay, Western Blot, Control, Mutagenesis

Journal: Life

Article Title: Identification of New, Functionally Relevant Mutations in the Coding Regions of the Human Fos and Jun Proto-Oncogenes in Rheumatoid Arthritis Synovial Tissue

doi: 10.3390/life11010005

Figure Lengend Snippet: Quantitation of MMP-1 mRNA expression in jun-/fos-transfected primary human fibroblast-like synoviocytes (FLS). ( A – C ) The graph shows the MMP-1 mRNA expression (detected by qPCR) in primary FLS derived from RA and OA patients (without detectable genetic variations in the jun and fos genes; n = 3 each, mean ± SD) following transfection of cjun and/or cfos expression vectors. ( A ) MMP-1 expression in FLS transfected with the indicated cfos variants. ( B ) MMP-1 expression in FLS transfected with the indicated cjun variants. ( C ) MMP-1 expression in FLS transfected with combinations of cfos and cjun variants. MMP-1 mRNA levels in fosWt- ( A ), junWt- ( B ), or fosWt/junWt-transfected cells ( C ) were set as 1. Statistical analyses: Mann–Whitney U-test, * p ≤ 0.05 vs. the respective controls; fos73: double mutant fos73/fos125.

Article Snippet: For cFos and cJun protein detection, SDS-PAGE (10%) was performed using 10 μL of each whole cell extract sample, followed by Western Blot analysis using the respective cFos and cJun antibodies from the TransAM AP-1 Family Kit (Active Motif, Rixensart, Belgium).

Techniques: Quantitation Assay, Expressing, Transfection, Derivative Assay, MANN-WHITNEY, Mutagenesis

Journal: Life

Article Title: Identification of New, Functionally Relevant Mutations in the Coding Regions of the Human Fos and Jun Proto-Oncogenes in Rheumatoid Arthritis Synovial Tissue

doi: 10.3390/life11010005

Figure Lengend Snippet: Quantitation of IL-6 mRNA expression in jun-/fos-transfected primary human FLS. ( A – C ) The graph shows the IL-6 mRNA expression (detected by qPCR) in primary FLS derived from RA and OA patients (without detectable genetic variations in the jun and fos genes; n = 3 each, mean ± SD) following transfection of cjun and/or cfos expression vectors. ( A ) IL-6 expression in FLS transfected with the indicated cfos variants. ( B ) IL-6 expression in FLS transfected with the indicated cjun variants. ( C ) IL-6 expression in FLS transfected with combinations of cfos and cjun variants. IL-6 mRNA levels in fosWt- ( A ), junWt- ( B ), or fosWt/junWt-transfected cells ( C ) were set as 1. Statistical analyses: Mann–Whitney U-test, * p ≤ 0.05 vs. the respective controls; fos73: double mutant fos73/fos125.

Article Snippet: For cFos and cJun protein detection, SDS-PAGE (10%) was performed using 10 μL of each whole cell extract sample, followed by Western Blot analysis using the respective cFos and cJun antibodies from the TransAM AP-1 Family Kit (Active Motif, Rixensart, Belgium).

Techniques: Quantitation Assay, Expressing, Transfection, Derivative Assay, MANN-WHITNEY, Mutagenesis

Journal: Life

Article Title: Identification of New, Functionally Relevant Mutations in the Coding Regions of the Human Fos and Jun Proto-Oncogenes in Rheumatoid Arthritis Synovial Tissue

doi: 10.3390/life11010005

Figure Lengend Snippet: Frequency analysis of mutations in GnomAD.

Article Snippet: For cFos and cJun protein detection, SDS-PAGE (10%) was performed using 10 μL of each whole cell extract sample, followed by Western Blot analysis using the respective cFos and cJun antibodies from the TransAM AP-1 Family Kit (Active Motif, Rixensart, Belgium).

Techniques: