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transcriptase polymerase chain reaction pcr  (New England Biolabs)
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A) Model for activation of transcription at the ORE by RBPJκ (adapted from ). RNAP, <t>RNA</t> <t>polymerase;</t> KMT2D, Lysine Methyltransferase 2D. B) Co-immunoprecipitation analysis of RBPJκ and p300 in cells expressing the indicated plasmids. C) MNRR1 overexpressing cells display an increase in the levels of H3K4me3, a marker of active transcription. D) MT-TA-CHCH (construct # 4, ) is sufficient to induce the activatory histone signature H3K4me3. E) Cells expressing wild type MNRR1 (WT) display an increased interaction between RBPJκ and H3K4me3 compared to cells expressing an empty vector (EV). F) MNRR1 in the presence of Sirt1 (that deacetylates it, thereby making it a superior transcription factor) displaces KMT2D and recruits p300 to RBPJκ. G) Cells expressing the deacetylated (K-R) but not the acetylmimetic (K-Q) mutant of MNRR1 display enhanced interaction between RBPJκ and p300.
Transcriptase Polymerase Chain Reaction Pcr, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A) Model for activation of transcription at the ORE by RBPJκ (adapted from ). RNAP, <t>RNA</t> <t>polymerase;</t> KMT2D, Lysine Methyltransferase 2D. B) Co-immunoprecipitation analysis of RBPJκ and p300 in cells expressing the indicated plasmids. C) MNRR1 overexpressing cells display an increase in the levels of H3K4me3, a marker of active transcription. D) MT-TA-CHCH (construct # 4, ) is sufficient to induce the activatory histone signature H3K4me3. E) Cells expressing wild type MNRR1 (WT) display an increased interaction between RBPJκ and H3K4me3 compared to cells expressing an empty vector (EV). F) MNRR1 in the presence of Sirt1 (that deacetylates it, thereby making it a superior transcription factor) displaces KMT2D and recruits p300 to RBPJκ. G) Cells expressing the deacetylated (K-R) but not the acetylmimetic (K-Q) mutant of MNRR1 display enhanced interaction between RBPJκ and p300.
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A) Model for activation of transcription at the ORE by RBPJκ (adapted from ). RNAP, <t>RNA</t> <t>polymerase;</t> KMT2D, Lysine Methyltransferase 2D. B) Co-immunoprecipitation analysis of RBPJκ and p300 in cells expressing the indicated plasmids. C) MNRR1 overexpressing cells display an increase in the levels of H3K4me3, a marker of active transcription. D) MT-TA-CHCH (construct # 4, ) is sufficient to induce the activatory histone signature H3K4me3. E) Cells expressing wild type MNRR1 (WT) display an increased interaction between RBPJκ and H3K4me3 compared to cells expressing an empty vector (EV). F) MNRR1 in the presence of Sirt1 (that deacetylates it, thereby making it a superior transcription factor) displaces KMT2D and recruits p300 to RBPJκ. G) Cells expressing the deacetylated (K-R) but not the acetylmimetic (K-Q) mutant of MNRR1 display enhanced interaction between RBPJκ and p300.
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Fig. 5 Representative flow cytometry and <t>ddPCR</t> analysis of LSCs and 3T3 cells prepared as the feeder layer. A and B Gating strategy of single-cell LSCs based on forward scatter (FSC) and side scatter (SSC) parameters. Unspecific staining was set to 1% of the acquired cells according to isotype antibody staining. C The majority of LSCs expressed p63α, D but few were positive for the ABCG2 stem cell-associated marker. E Residual 3T3 feeder cells in LSC expanded culture were below the acceptance limit. F 1D plot for two sets of analyses on ddPCR analysis of sample 3 (A1 – D1) and sample 4 (E1 – H1) with mouse assay. G4 is analysis of mouse positive control (DNA from pure 3T3 culture), H11 is negative control (NTC) and H12 is test for possible cross-reactivity with human DNA (human positive control). G Analysis of sample 3 (A3) and sample 4 (B3) with the human assay. H4 is analysis of human positive control (DNA from human whole blood), A11 is NTC and A12 is test for possible cross-reactivity with mouse DNA (mouse positive control)
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Fig. 5 Representative flow cytometry and <t>ddPCR</t> analysis of LSCs and 3T3 cells prepared as the feeder layer. A and B Gating strategy of single-cell LSCs based on forward scatter (FSC) and side scatter (SSC) parameters. Unspecific staining was set to 1% of the acquired cells according to isotype antibody staining. C The majority of LSCs expressed p63α, D but few were positive for the ABCG2 stem cell-associated marker. E Residual 3T3 feeder cells in LSC expanded culture were below the acceptance limit. F 1D plot for two sets of analyses on ddPCR analysis of sample 3 (A1 – D1) and sample 4 (E1 – H1) with mouse assay. G4 is analysis of mouse positive control (DNA from pure 3T3 culture), H11 is negative control (NTC) and H12 is test for possible cross-reactivity with human DNA (human positive control). G Analysis of sample 3 (A3) and sample 4 (B3) with the human assay. H4 is analysis of human positive control (DNA from human whole blood), A11 is NTC and A12 is test for possible cross-reactivity with mouse DNA (mouse positive control)
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Fig. 5 Representative flow cytometry and <t>ddPCR</t> analysis of LSCs and 3T3 cells prepared as the feeder layer. A and B Gating strategy of single-cell LSCs based on forward scatter (FSC) and side scatter (SSC) parameters. Unspecific staining was set to 1% of the acquired cells according to isotype antibody staining. C The majority of LSCs expressed p63α, D but few were positive for the ABCG2 stem cell-associated marker. E Residual 3T3 feeder cells in LSC expanded culture were below the acceptance limit. F 1D plot for two sets of analyses on ddPCR analysis of sample 3 (A1 – D1) and sample 4 (E1 – H1) with mouse assay. G4 is analysis of mouse positive control (DNA from pure 3T3 culture), H11 is negative control (NTC) and H12 is test for possible cross-reactivity with human DNA (human positive control). G Analysis of sample 3 (A3) and sample 4 (B3) with the human assay. H4 is analysis of human positive control (DNA from human whole blood), A11 is NTC and A12 is test for possible cross-reactivity with mouse DNA (mouse positive control)
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A) Model for activation of transcription at the ORE by RBPJκ (adapted from ). RNAP, RNA polymerase; KMT2D, Lysine Methyltransferase 2D. B) Co-immunoprecipitation analysis of RBPJκ and p300 in cells expressing the indicated plasmids. C) MNRR1 overexpressing cells display an increase in the levels of H3K4me3, a marker of active transcription. D) MT-TA-CHCH (construct # 4, ) is sufficient to induce the activatory histone signature H3K4me3. E) Cells expressing wild type MNRR1 (WT) display an increased interaction between RBPJκ and H3K4me3 compared to cells expressing an empty vector (EV). F) MNRR1 in the presence of Sirt1 (that deacetylates it, thereby making it a superior transcription factor) displaces KMT2D and recruits p300 to RBPJκ. G) Cells expressing the deacetylated (K-R) but not the acetylmimetic (K-Q) mutant of MNRR1 display enhanced interaction between RBPJκ and p300.

Journal: bioRxiv

Article Title: Transcriptional activation by MNRR1 is effected by recruiting p300 and can be induced by minimal peptides

doi: 10.1101/2025.08.25.672219

Figure Lengend Snippet: A) Model for activation of transcription at the ORE by RBPJκ (adapted from ). RNAP, RNA polymerase; KMT2D, Lysine Methyltransferase 2D. B) Co-immunoprecipitation analysis of RBPJκ and p300 in cells expressing the indicated plasmids. C) MNRR1 overexpressing cells display an increase in the levels of H3K4me3, a marker of active transcription. D) MT-TA-CHCH (construct # 4, ) is sufficient to induce the activatory histone signature H3K4me3. E) Cells expressing wild type MNRR1 (WT) display an increased interaction between RBPJκ and H3K4me3 compared to cells expressing an empty vector (EV). F) MNRR1 in the presence of Sirt1 (that deacetylates it, thereby making it a superior transcription factor) displaces KMT2D and recruits p300 to RBPJκ. G) Cells expressing the deacetylated (K-R) but not the acetylmimetic (K-Q) mutant of MNRR1 display enhanced interaction between RBPJκ and p300.

Article Snippet: Complementary DNA (cDNA) was generated by reverse transcriptase polymerase chain reaction (PCR) using the ProtoScript ® II First Strand cDNA Synthesis Kit (New England Biolabs, Ipswich, MA, USA).

Techniques: Activation Assay, Immunoprecipitation, Expressing, Marker, Construct, Plasmid Preparation, Mutagenesis

Fig. 5 Representative flow cytometry and ddPCR analysis of LSCs and 3T3 cells prepared as the feeder layer. A and B Gating strategy of single-cell LSCs based on forward scatter (FSC) and side scatter (SSC) parameters. Unspecific staining was set to 1% of the acquired cells according to isotype antibody staining. C The majority of LSCs expressed p63α, D but few were positive for the ABCG2 stem cell-associated marker. E Residual 3T3 feeder cells in LSC expanded culture were below the acceptance limit. F 1D plot for two sets of analyses on ddPCR analysis of sample 3 (A1 – D1) and sample 4 (E1 – H1) with mouse assay. G4 is analysis of mouse positive control (DNA from pure 3T3 culture), H11 is negative control (NTC) and H12 is test for possible cross-reactivity with human DNA (human positive control). G Analysis of sample 3 (A3) and sample 4 (B3) with the human assay. H4 is analysis of human positive control (DNA from human whole blood), A11 is NTC and A12 is test for possible cross-reactivity with mouse DNA (mouse positive control)

Journal: Stem cell research & therapy

Article Title: New characterization and safety evaluation of human limbal stem cells used in clinical application: fidelity of mitotic process and mitotic spindle morphologies.

doi: 10.1186/s13287-023-03586-z

Figure Lengend Snippet: Fig. 5 Representative flow cytometry and ddPCR analysis of LSCs and 3T3 cells prepared as the feeder layer. A and B Gating strategy of single-cell LSCs based on forward scatter (FSC) and side scatter (SSC) parameters. Unspecific staining was set to 1% of the acquired cells according to isotype antibody staining. C The majority of LSCs expressed p63α, D but few were positive for the ABCG2 stem cell-associated marker. E Residual 3T3 feeder cells in LSC expanded culture were below the acceptance limit. F 1D plot for two sets of analyses on ddPCR analysis of sample 3 (A1 – D1) and sample 4 (E1 – H1) with mouse assay. G4 is analysis of mouse positive control (DNA from pure 3T3 culture), H11 is negative control (NTC) and H12 is test for possible cross-reactivity with human DNA (human positive control). G Analysis of sample 3 (A3) and sample 4 (B3) with the human assay. H4 is analysis of human positive control (DNA from human whole blood), A11 is NTC and A12 is test for possible cross-reactivity with mouse DNA (mouse positive control)

Article Snippet: Each 20 μL droplet digital polymerase chain reaction (ddPCR) consisted of 10 μL of ddPCR Supermix for Probes (Bio-Rad, USA), 1 μL of Hind III (New England Biolabs, USA) and 4 μL of DNA.

Techniques: Flow Cytometry, Staining, Marker, Mouse Assay, Positive Control, Negative Control