rabbit anti-mtap polyclonal antibody  (Cell Signaling Technology Inc)

Journal: medRxiv

Article Title: Enhancer-targeting CRISPR screens at coronary artery disease loci suggest shared mechanisms of disease risk

doi: 10.1101/2025.08.28.25334684

Figure Lengend Snippet: A) Design of SMC in vitro DC-TAP-seq targeting screen of CAD enhancers at 9p21.3. B) Details regarding CAD GWAS enhancer identification, gRNA design for CRISPRi epigenome editing, and screen results. C) Graphical representation of guide RNAs (gRNAs, protospacers) transduced into CRISPRi-HCASMC by lentiviral infection. D) Violin plot showing effect of gRNA directed gene knockdown and resulting effects on gene expression. Experimental conditions were gRNA directed to CAD GWAS enhancers, targeting of TSS for 9p21.3 experimental genes, and targeting of TSS for known CAD causal genes PDGFD and SMAD3 . E) Volcano plot showing effect on gene expression of gRNA targeted enhancers identified at 9p21.3, graphed as log fold change versus effect size. Grey dots represent genes not affected by enhancer knockdown, red dots and gene symbols represent genes showing significant effect toward gRNA targeting, average of multiple guides targeted to specific sequences in the 9p21.3 enhancers. F) Genome browser image showing long range chromatin organization at this locus, H3K27ac histone modification from ChIPseq, bulk ATACseq, and HiC loops all mapped in telo-HCASMC. Also, shown are genes encoded in this locus, and the E2G links identified by DC-TAP-seq. G, H) CRISPRi epigenome editing in CRISPRi-HCASMC transduced with lentivirus control gRNAs, no gRNAs, CDKN2A transcription start site (TSS), CDKN2B TSS, and sgRNAs for enhancers expected to be positive or negative. I) A similar PCR quantitation of MTAP expression was performed with CRISPRi-HCASMC transduced with lentivirus control gRNAs, no gRNAs, and gRNAs targeting the TSS of CAD Twist1 as a positive control, and sgRNAs for enhancers expected to be positive or negative. J) Enhancer control of ANRIL expression was investigated with gRNAs targeting E2, E4, and E6. Experiments were conducted with CDKN2A and CDKN2B TSS gRNAs to investigate gene crosstalk.

Article Snippet: Gene expression was assessed using TaqMan qPCR probes (Thermo Fisher) for CDKN2A (HS99999189_M1), CDKN2B (Hs00793225_m1), CDKN2B-AS1 (Hs03300540_m1), MTAP (HS00559618_M1), MICA (Hs07292198_gH), SBF2 (Hs00959709_g1), TP53INP3 (Hs00894008_g1), RAB23 (Hs00212407_m1), PPP1R18 (Hs00292978_m1), CKB (HS00176484_M1) according to the manufacturer’s instructions on a ViiA7 Real-Time PCR system (Applied Biosystems, Foster City, CA).

Techniques: In Vitro, Infection, Knockdown, Gene Expression, Modification, Transduction, Control, Quantitation Assay, Expressing, Positive Control

Journal: medRxiv

Article Title: Enhancer-targeting CRISPR screens at coronary artery disease loci suggest shared mechanisms of disease risk

doi: 10.1101/2025.08.28.25334684

Figure Lengend Snippet: A, B) 9p21.3 V2E2G individual gRNA level DC-TAP-seq data. C-F) CRISPRa validations for V2E2G for ANRIL , CDKN2B, CDKN2A, and MTAP .

Article Snippet: Gene expression was assessed using TaqMan qPCR probes (Thermo Fisher) for CDKN2A (HS99999189_M1), CDKN2B (Hs00793225_m1), CDKN2B-AS1 (Hs03300540_m1), MTAP (HS00559618_M1), MICA (Hs07292198_gH), SBF2 (Hs00959709_g1), TP53INP3 (Hs00894008_g1), RAB23 (Hs00212407_m1), PPP1R18 (Hs00292978_m1), CKB (HS00176484_M1) according to the manufacturer’s instructions on a ViiA7 Real-Time PCR system (Applied Biosystems, Foster City, CA).

Techniques:

Journal: iScience

Article Title: The PRMT5-splicing axis is a critical oncogenic vulnerability that regulates detained intron splicing

doi: 10.1016/j.isci.2025.112965

Figure Lengend Snippet: PRMT5 regulates a conserved set of DIs across multiple human cell lines (A) Human cell lines used for AS analysis, including cancer or tissue of origin, driver mutations or construct, and MTAP status. (B) Proportion of each AS event type that show significantly altered levels in the indicated cell line in response to 3-day treatment with 10 nM JNJ-64619178 (PRMT5i) compared to vehicle control (p adj < 0.05). The total number of significant AS events for that cell line is indicated to the right of each bar. (C) Relative expression of individual DIs across all human cell lines after 3-day vehicle or PRMT5i treatment. Each column represents a specific DI that is significantly up- or down-regulated by PRMT5i-treatment in at least one cell line. Coloring represents Z score normalized intron counts, where red indicates higher relative expression and blue indicates lower relative expression. (D) Similarity matrix of the indicated AS event significantly altered by PRMT5i treatment (p adj < 0.05). Each square represents the similarity between the two intersecting cell lines, with coloring indicating percent similarity (dice similarity score) and square size indicating the number of similar events. (E) Bar charts of DI conservation across 7 human cell lines. Bar length represents the number of significant PRMT5i-upregulated DIs (p adj < 0.05, log 2 FC > 0) for each cell line and the color denotes the number of cell lines in which a given DI is conserved across. The total number of significant DIs in each line is indicated to the right of each bar. (F) Enriched GO terms from the KW Biological Process gene set for PRMT5i-upregulated DIs in each cell line (p adj < 0.05, log 2 FC > 0). Terms are displayed if they are significant in at least one cell line (FDR < 0.01). Color represents enrichment score, while size inversely correlates with significance value.

Article Snippet: Rabbit anti-MTAP Polyclonal Antibody , Cell Signaling Technology , Cat#4158; RRID: AB_1904054.

Techniques: Construct, Control, Expressing

Journal: iScience

Article Title: The PRMT5-splicing axis is a critical oncogenic vulnerability that regulates detained intron splicing

doi: 10.1016/j.isci.2025.112965

Figure Lengend Snippet: PRMT5 regulates a conserved set of DIs in multiple mouse cell lines (A) Summary of mouse cell lines used for AS analysis, including cancer type, driver mutations, PRMT5i used, and MTAP status. (B) Proportion of each AS event type that is significantly different in indicated cell line after 3-day PRMT5i treatment compared to vehicle control (p adj < 0.05). Total number of significant AS events is indicated above the bar. (C) Relative expression of individual DIs across all mouse cell lines after 3-day vehicle or PRMT5i treatment. Each column represents a specific DI that is significantly upregulated or downregulated by PRMT5i-treatment in at least one cell line. Colors represent column Z score normalized intron counts, where red indicates higher relative expression and blue indicates lower relative expression. (D) Similarity matrix of the indicated class of AS event that was significantly altered by PRMT5i treatment. Each square represents the similarity between the two intersecting murine cell lines. Square coloring indicates percent similarity (dice similarity score), and square size is proportional to the number of similar events. (E) Bar charts of DI conservation across all mouse cell lines. Each bar represents the number of PRMT5i-upregulated DIs (p adj < 0.05, log 2 FC > 0) in each cell line and the color corresponds to the number of cell lines in which a given DI is conserved across. Total number of significant DIs in each line is indicated to the right of the bar. (F) Enriched GO terms from the KW Biological Process gene set for PRMT5i-upregualted DIs in each cell line (p adj < 0.05, log 2 FC > 0). Terms are displayed if they are significant in at least one cell line (FDR < 0.01). Color represents enrichment score, while size inversely correlates with significance value.

Article Snippet: Rabbit anti-MTAP Polyclonal Antibody , Cell Signaling Technology , Cat#4158; RRID: AB_1904054.

Techniques: Control, Expressing