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anti mef2b  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology anti mef2b
    Anti Mef2b, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Article Title: Direct reprogramming of fibroblasts into skeletal muscle progenitor cells by transcription factors enriched in undifferentiated subpopulation of satellite cells
    Article Snippet: These plasmids were recombined with the CSIV-TRE-RfA-CMV-KT vector by LR reaction (Thermo Fisher Scientific).

    Article Title: Disruption of chaperone-mediated autophagy-dependent degradation of MEF2A by oxidative stress-induced lysosome destabilization
    Article Snippet: Anti-MEF2A (sc-313), anti-MEF2B (sc-101097), anti-MEF2C (sc-365862), anti-MEF2D (sc-271153), anti-ACTB (β-actin, sc-47778) and goat anti-mouse (sc-2005) or rabbit (sc-2004) IgG-HRP were purchased from Santa Cruz Biotechnology.

    Article Title: Unique immune cell coactivators specify locus control region function and cell stage
    Article Snippet: Anti-MEF2B, mouse monoclonal , Santa Cruz Biotechnology , Cat# sc-517433; RRID: NA.

    Article Title: Contribution of Myocyte Enhancer Factor 2 Family Transcription Factors to BZLF1 Expression in Epstein-Barr Virus Reactivation from Latency
    Article Snippet: Anti-MEF2A (TA303858) antibody was obtained from OriGene Technologies, and anti-MEF2B (sc-98594) and anti-MEF2D (sc-271153) antibodies were from Santa Cruz Biotechnology.

    Article Title: DNA methylation remodeling in temozolomide resistant recurrent glioblastoma: comparing epigenetic dynamics in vitro and in vivo.
    Article Snippet: We purchased the following antibodies: antiα-tubulin (Abcam, Cambridge), anti-MGMT (Santa Cruz Biotechnology, Texas), anti-MEF2B (Santa Cruz Biotechnology), anti-MAPK6 (Santa Cruz Biotechnology).

    Article Title: Disruption of chaperone-mediated autophagy-dependent degradation of MEF2A by oxidative stress-induced lysosome destabilization
    Article Snippet: Antibodies and reagents Anti-MEF2A (sc-313), anti-MEF2B (sc-101097), anti-MEF2C (sc-365862), anti-MEF2D (sc-271153), anti-ACTB (β-actin, sc-47778) and goat anti-mouse (sc-2005) or rabbit (sc-2004) IgG-HRP were purchased from Santa Cruz Biotechnology.

    Article Title: Characterization of a Cardiac-specific Enhancer, Which Directs α-Cardiac Actin Gene Transcription in the Mouse Adult Heart
    Article Snippet: B, the complexes formed (lane 1) were also probed by preincubation with a preimmune serum (lane 2), with a generic MEF2 antibody obtained from Santa Cruz (lane 3), with monospecific anti-MEF2A (lane 4), anti-MEF2B (lane 5), anti-MEF2D (lane 6), and with antibodies to mHox (lane 7) and Oct1 (lane 8). at W A SH B U R N U N IV E R SIT Y on M arch 4, 2015 http://w w w .jbc.org/ D ow nloaded from around this site suggests that in this case MEF2D may participate in a cardiac-specific regulatory complex.

    Article Title: DNA methylation remodeling in temozolomide resistant recurrent glioblastoma: comparing epigenetic dynamics in vitro and in vivo
    Article Snippet: We purchased the following antibodies: anti-α-tubulin (Abcam, Cambridge), anti-MGMT (Santa Cruz Biotechnology, Texas), anti-MEF2B (Santa Cruz Biotechnology), anti-MAPK6 (Santa Cruz Biotechnology).



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    Figure 3. Activation of MYC by ternary complex–bound modules of the BCL6 super-enhancer. A, Schematic diagram of the normal BCL6 locus and parsimonious models for structural rearrangements involving the MYC and BCL6 locus in three DLBCL cell lines based on optical mapping and 4C-seq data. Red lines indicate intra- or inter-chromosomal fusions. See Supplementary Fig. S3A–S3C for details. B, CRISPRi sgRNA depletion or enrichment (log2 fold change) for BCL6 distal super-enhancer regions in the MYC::BCL6-LCR+ cell lines SU-DHL-4 and WSU-DLCL2. Also shown is ChIP-seq signal for H3K27ac and ternary complex TFs <t>MEF2B</t> and OCT2 in SU-DHL-4 (scale: fragments per million mapped fragments). BCL6 locus super-enhancer regions SE1 and SE3 are shown as previously defined (15); region SE1 is equivalent to BCL6-LCR. C, Plot of NFR-level sgRNA depletion or enrichment (MAGeCK RRA log2 fold change, y-axis) for targeted intervals in BCL6 distal super-enhancers in six GCB-DLBCL cell lines. Circles are sized proportionally to −log2(FDR), with neg|FDR used for log2FC <0 and pos|FDR used for log2FC >0. Filled circles represent neg|FDR <0.01. The three MYC::BCL6-LCR+ cell lines are indicated in the legend with a red box. D, Change in viable cellularity for dox-inducible KRAB-dCas9–expressing MYC::BCL6-LCR+ DLBCL cell lines expressing the two indicated sgRNA lentivectors. Cells were transduced with both vectors, sorted for dual tagBFP- and tagRFP-expressing cells, and then grown with or without dox for 7 days. The Y-axis shows the ratio of CellTiter-Glo signal for dox treated/untreated cells (two-tailed Student t test of n = 3 biological replicates per condition for dual-enhancer–targeting sgRNA-transduced vs. dual-control sgRNA-transduced; error bars are 95% CI). E, Differential expression of MYC and BCL6 (qRT-PCR) for populations of MYC::BCL6-LCR+ DLBCL cell lines expressing dox-inducible KRAB-dCas9 and two sgRNA vectors as in (D) (two-tailed Student t test of n = 3 technical replicates per condition for dual-enhancer–targeting sgRNA-transduced vs. dual-control sgRNA-transduced, color-coded by measured transcript per legend; error bars are 95% CI). F, Differential expression of MYC and BCL6 (qRT-PCR) in MYC::BCL6-LCR+ DLBCL cell lines express- ing dox-inducible KRAB-dCas9 after transduction with indicated sgRNAs targeting MEF2B, POU2F2, or POU2AF1 (error bars are the SD of n = 5 technical replicates per sgRNA; bars are color-coded by measured transcript per legend). ***, P < 0.001. FC, fold change; neg ctrl, negative control; pos, positive.
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    Fig. 7 | Stabilization of <t>MEF2B</t> C-terminal isoform-switch mutants by dis- sociation from the CUL3/KLHL12 complex. a Detection of MEF2B/CUL3/KLHL12 interaction by Flag (MEF2B) immunoprecipitation (IP) using nuclear extracts of SUDHL10 cells expressing Flag-HA-MEF2B-WT-isoA, -isoB or isoform-switch mutants. Lamin B1 was used as loading control. Quantifications were normalized to the immunoprecipitated MEF2B (HA). Fold changes were calculated relative to the MEF2B WT-isoA sample. b MEF2B IP followed by immunoblotting (IB) using nuclear extracts of isogenic OCI-Ly7 clones including controls (Neutral) and cells expres- sing MEF2B isoform-switch (P256fs and P273fs) mutants. IgG IP was used as nega- tive control. Quantifications were normalized to the immunoprecipitated MEF2B. Fold changes were calculated relative to the neutral clones. Samples are the same as in Fig. 5e. c Ubiquitination assay upon Flag (MEF2B) IP followed by IB using total extracts of HEK293T cells co-transfected with expression vectors for Flag-HA- MEF2B-WT-isoA, -isoB or isoform-switch mutants (P243fs, L269fs, and P273fs), and
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    Fig. 7 | Stabilization of <t>MEF2B</t> C-terminal isoform-switch mutants by dis- sociation from the CUL3/KLHL12 complex. a Detection of MEF2B/CUL3/KLHL12 interaction by Flag (MEF2B) immunoprecipitation (IP) using nuclear extracts of SUDHL10 cells expressing Flag-HA-MEF2B-WT-isoA, -isoB or isoform-switch mutants. Lamin B1 was used as loading control. Quantifications were normalized to the immunoprecipitated MEF2B (HA). Fold changes were calculated relative to the MEF2B WT-isoA sample. b MEF2B IP followed by immunoblotting (IB) using nuclear extracts of isogenic OCI-Ly7 clones including controls (Neutral) and cells expres- sing MEF2B isoform-switch (P256fs and P273fs) mutants. IgG IP was used as nega- tive control. Quantifications were normalized to the immunoprecipitated MEF2B. Fold changes were calculated relative to the neutral clones. Samples are the same as in Fig. 5e. c Ubiquitination assay upon Flag (MEF2B) IP followed by IB using total extracts of HEK293T cells co-transfected with expression vectors for Flag-HA- MEF2B-WT-isoA, -isoB or isoform-switch mutants (P243fs, L269fs, and P273fs), and
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    Fig. 7 | Stabilization of <t>MEF2B</t> C-terminal isoform-switch mutants by dis- sociation from the CUL3/KLHL12 complex. a Detection of MEF2B/CUL3/KLHL12 interaction by Flag (MEF2B) immunoprecipitation (IP) using nuclear extracts of SUDHL10 cells expressing Flag-HA-MEF2B-WT-isoA, -isoB or isoform-switch mutants. Lamin B1 was used as loading control. Quantifications were normalized to the immunoprecipitated MEF2B (HA). Fold changes were calculated relative to the MEF2B WT-isoA sample. b MEF2B IP followed by immunoblotting (IB) using nuclear extracts of isogenic OCI-Ly7 clones including controls (Neutral) and cells expres- sing MEF2B isoform-switch (P256fs and P273fs) mutants. IgG IP was used as nega- tive control. Quantifications were normalized to the immunoprecipitated MEF2B. Fold changes were calculated relative to the neutral clones. Samples are the same as in Fig. 5e. c Ubiquitination assay upon Flag (MEF2B) IP followed by IB using total extracts of HEK293T cells co-transfected with expression vectors for Flag-HA- MEF2B-WT-isoA, -isoB or isoform-switch mutants (P243fs, L269fs, and P273fs), and
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    Figure 3. Activation of MYC by ternary complex–bound modules of the BCL6 super-enhancer. A, Schematic diagram of the normal BCL6 locus and parsimonious models for structural rearrangements involving the MYC and BCL6 locus in three DLBCL cell lines based on optical mapping and 4C-seq data. Red lines indicate intra- or inter-chromosomal fusions. See Supplementary Fig. S3A–S3C for details. B, CRISPRi sgRNA depletion or enrichment (log2 fold change) for BCL6 distal super-enhancer regions in the MYC::BCL6-LCR+ cell lines SU-DHL-4 and WSU-DLCL2. Also shown is ChIP-seq signal for H3K27ac and ternary complex TFs MEF2B and OCT2 in SU-DHL-4 (scale: fragments per million mapped fragments). BCL6 locus super-enhancer regions SE1 and SE3 are shown as previously defined (15); region SE1 is equivalent to BCL6-LCR. C, Plot of NFR-level sgRNA depletion or enrichment (MAGeCK RRA log2 fold change, y-axis) for targeted intervals in BCL6 distal super-enhancers in six GCB-DLBCL cell lines. Circles are sized proportionally to −log2(FDR), with neg|FDR used for log2FC <0 and pos|FDR used for log2FC >0. Filled circles represent neg|FDR <0.01. The three MYC::BCL6-LCR+ cell lines are indicated in the legend with a red box. D, Change in viable cellularity for dox-inducible KRAB-dCas9–expressing MYC::BCL6-LCR+ DLBCL cell lines expressing the two indicated sgRNA lentivectors. Cells were transduced with both vectors, sorted for dual tagBFP- and tagRFP-expressing cells, and then grown with or without dox for 7 days. The Y-axis shows the ratio of CellTiter-Glo signal for dox treated/untreated cells (two-tailed Student t test of n = 3 biological replicates per condition for dual-enhancer–targeting sgRNA-transduced vs. dual-control sgRNA-transduced; error bars are 95% CI). E, Differential expression of MYC and BCL6 (qRT-PCR) for populations of MYC::BCL6-LCR+ DLBCL cell lines expressing dox-inducible KRAB-dCas9 and two sgRNA vectors as in (D) (two-tailed Student t test of n = 3 technical replicates per condition for dual-enhancer–targeting sgRNA-transduced vs. dual-control sgRNA-transduced, color-coded by measured transcript per legend; error bars are 95% CI). F, Differential expression of MYC and BCL6 (qRT-PCR) in MYC::BCL6-LCR+ DLBCL cell lines express- ing dox-inducible KRAB-dCas9 after transduction with indicated sgRNAs targeting MEF2B, POU2F2, or POU2AF1 (error bars are the SD of n = 5 technical replicates per sgRNA; bars are color-coded by measured transcript per legend). ***, P < 0.001. FC, fold change; neg ctrl, negative control; pos, positive.

    Journal: Blood Cancer Discovery

    Article Title: Selective Enhancer Dependencies in MYC-Intact and MYC-Rearranged Germinal Center B-cell Diffuse Large B-cell Lymphoma

    doi: 10.1158/2643-3230.bcd-24-0126

    Figure Lengend Snippet: Figure 3. Activation of MYC by ternary complex–bound modules of the BCL6 super-enhancer. A, Schematic diagram of the normal BCL6 locus and parsimonious models for structural rearrangements involving the MYC and BCL6 locus in three DLBCL cell lines based on optical mapping and 4C-seq data. Red lines indicate intra- or inter-chromosomal fusions. See Supplementary Fig. S3A–S3C for details. B, CRISPRi sgRNA depletion or enrichment (log2 fold change) for BCL6 distal super-enhancer regions in the MYC::BCL6-LCR+ cell lines SU-DHL-4 and WSU-DLCL2. Also shown is ChIP-seq signal for H3K27ac and ternary complex TFs MEF2B and OCT2 in SU-DHL-4 (scale: fragments per million mapped fragments). BCL6 locus super-enhancer regions SE1 and SE3 are shown as previously defined (15); region SE1 is equivalent to BCL6-LCR. C, Plot of NFR-level sgRNA depletion or enrichment (MAGeCK RRA log2 fold change, y-axis) for targeted intervals in BCL6 distal super-enhancers in six GCB-DLBCL cell lines. Circles are sized proportionally to −log2(FDR), with neg|FDR used for log2FC <0 and pos|FDR used for log2FC >0. Filled circles represent neg|FDR <0.01. The three MYC::BCL6-LCR+ cell lines are indicated in the legend with a red box. D, Change in viable cellularity for dox-inducible KRAB-dCas9–expressing MYC::BCL6-LCR+ DLBCL cell lines expressing the two indicated sgRNA lentivectors. Cells were transduced with both vectors, sorted for dual tagBFP- and tagRFP-expressing cells, and then grown with or without dox for 7 days. The Y-axis shows the ratio of CellTiter-Glo signal for dox treated/untreated cells (two-tailed Student t test of n = 3 biological replicates per condition for dual-enhancer–targeting sgRNA-transduced vs. dual-control sgRNA-transduced; error bars are 95% CI). E, Differential expression of MYC and BCL6 (qRT-PCR) for populations of MYC::BCL6-LCR+ DLBCL cell lines expressing dox-inducible KRAB-dCas9 and two sgRNA vectors as in (D) (two-tailed Student t test of n = 3 technical replicates per condition for dual-enhancer–targeting sgRNA-transduced vs. dual-control sgRNA-transduced, color-coded by measured transcript per legend; error bars are 95% CI). F, Differential expression of MYC and BCL6 (qRT-PCR) in MYC::BCL6-LCR+ DLBCL cell lines express- ing dox-inducible KRAB-dCas9 after transduction with indicated sgRNAs targeting MEF2B, POU2F2, or POU2AF1 (error bars are the SD of n = 5 technical replicates per sgRNA; bars are color-coded by measured transcript per legend). ***, P < 0.001. FC, fold change; neg ctrl, negative control; pos, positive.

    Article Snippet: The following primary antibodies were used: MEF2B (Atlas Antibodies #HPA004734, RRID: AB_10963939), EBF (Santa Cruz Biotechnology #sc-137065, RRID: AB_2246405), E2A (Santa Cruz Biotechnology #sc-349, RRID: AB_675504), Bob1 (Santa Cruz Biotechnology #sc-955, RRID: AB_2166917), Oct-2 (Santa Cruz Biotechnology #sc-233, RRID: AB_2167205), Bcl-6 (Santa Cruz Biotechnology #sc-7388, RRID: AB_2063455), c-Myc (Cell Signaling Technology #5605, RRID: AB_1903938), CTCF (Cell Signaling Technology #3418, RRID: AB_2086791), c-Rel (Cell Signaling Technology #12707, RRID: AB_2721030), phosphoSer536-RelA (Cell Signaling Technology #3033, RRID: AB_331284), RelA (Cell Signaling Technology #8242, RRID: AB_10859369), NF-κB1/p50 (Cell Signaling Technology #12540, RRID: AB_2687614), NF-κB2/p52 (Cell Signaling Technology #3017, RRID: AB_10697356), and TBP (Abcam #ab818, RRID: AB_306337).

    Techniques: Activation Assay, ChIP-sequencing, Expressing, Transduction, Two Tailed Test, Control, Quantitative Proteomics, Quantitative RT-PCR, Negative Control

    Figure 5. The GME-1 MYC enhancer shows selective transactivation in MYC-intact cell lines and is directly regulated by the OCT2 ternary com- plex. A, Detail of the GME-1 enhancer with ATAC-seq, H3K27 ChIP-seq, and ChIP-seq for OCT2, OCA-B, MEF2B, and p300 in the indicated cell lines. OCT2 BioChIP in BJAB cells is from Hodson and colleagues (39) and represents binding of a transgenically expressed biotinylated OCT2 protein, whereas all other ChIP-seq datasets profile endogenous proteins. Inset at bottom shows further detail of the Phastcons conserved element track for sub-region 195B (see Supplementary Fig. S5E for further detail), with the positions of the conserved OCT2 motif and the genomic duplica- tion identified in the SU-DHL-5 cell line indicated. B, Differential expression of MYC and BCL6 (qRT-PCR) in GME-1–dependent DLBCL cell lines expressing dox-inducible KRAB-dCas9 after transduction with indicated sgRNAs targeting MEF2B, POU2F2, and POU2AF1 (error bars are the SD of five technical replicates per sgRNA; bars are color-coded by measured transcript per legend). C, Transcriptional reporter assays conducted in MYC-intact and MYC::BCL6-LCR+ DLBCL cell lines with conserved subregions of essential MYC and BCL6 NFRs cloned into the STARR-seq_Ori lu- ciferase vector. See Supplementary Fig. S5E–S5H for details of the indicated regions. The signal ratio and statistical comparison for MYC enhancer 195B vs. each BCL6 enhancer is indicated for each cell line (two-tailed Student t test of biological replicates (n = 3); error bars are 95% confidence interval). D, Transcriptional reporter assays in MYC-intact cell lines for constructs bearing the reference NFR 195B enhancer region or the same region with deletion of the OCT2 motif (“195B_MYC_OCT2Del”) indicated in (A) [two-tailed Student t test of biological replicates (n = 3); error bars are 95% CI]. E, Transcriptional reporter assays conducted in HT cells expressing inducible KRAB-dCas9 and transduced with the indicated sgRNAs. Following puromycin selection, cells were electroporated with the indicated constructs and induced with dox for 48 hours prior to Dual-Glo lucifer- ase assay [two-tailed Student t test of biological replicates (n = 3); error bars are 95% CI]. F, Transcriptional reporter assays in MYC-intact cell lines for constructs bearing the reference NFR 195B enhancer region or a clone of the same region from the SU-DHL-5 cell line bearing a partial tandem duplication (“195B_MYC_Dup”) as indicated in (A) [two-tailed Student t test of biological replicates (n = 3); error bars are 95% CI]. **, P < 0.01; ***, P < 0.001; ns, not significant; P ≥ 0.05. FC, fold change; neg ctrl, negative control.

    Journal: Blood Cancer Discovery

    Article Title: Selective Enhancer Dependencies in MYC-Intact and MYC-Rearranged Germinal Center B-cell Diffuse Large B-cell Lymphoma

    doi: 10.1158/2643-3230.bcd-24-0126

    Figure Lengend Snippet: Figure 5. The GME-1 MYC enhancer shows selective transactivation in MYC-intact cell lines and is directly regulated by the OCT2 ternary com- plex. A, Detail of the GME-1 enhancer with ATAC-seq, H3K27 ChIP-seq, and ChIP-seq for OCT2, OCA-B, MEF2B, and p300 in the indicated cell lines. OCT2 BioChIP in BJAB cells is from Hodson and colleagues (39) and represents binding of a transgenically expressed biotinylated OCT2 protein, whereas all other ChIP-seq datasets profile endogenous proteins. Inset at bottom shows further detail of the Phastcons conserved element track for sub-region 195B (see Supplementary Fig. S5E for further detail), with the positions of the conserved OCT2 motif and the genomic duplica- tion identified in the SU-DHL-5 cell line indicated. B, Differential expression of MYC and BCL6 (qRT-PCR) in GME-1–dependent DLBCL cell lines expressing dox-inducible KRAB-dCas9 after transduction with indicated sgRNAs targeting MEF2B, POU2F2, and POU2AF1 (error bars are the SD of five technical replicates per sgRNA; bars are color-coded by measured transcript per legend). C, Transcriptional reporter assays conducted in MYC-intact and MYC::BCL6-LCR+ DLBCL cell lines with conserved subregions of essential MYC and BCL6 NFRs cloned into the STARR-seq_Ori lu- ciferase vector. See Supplementary Fig. S5E–S5H for details of the indicated regions. The signal ratio and statistical comparison for MYC enhancer 195B vs. each BCL6 enhancer is indicated for each cell line (two-tailed Student t test of biological replicates (n = 3); error bars are 95% confidence interval). D, Transcriptional reporter assays in MYC-intact cell lines for constructs bearing the reference NFR 195B enhancer region or the same region with deletion of the OCT2 motif (“195B_MYC_OCT2Del”) indicated in (A) [two-tailed Student t test of biological replicates (n = 3); error bars are 95% CI]. E, Transcriptional reporter assays conducted in HT cells expressing inducible KRAB-dCas9 and transduced with the indicated sgRNAs. Following puromycin selection, cells were electroporated with the indicated constructs and induced with dox for 48 hours prior to Dual-Glo lucifer- ase assay [two-tailed Student t test of biological replicates (n = 3); error bars are 95% CI]. F, Transcriptional reporter assays in MYC-intact cell lines for constructs bearing the reference NFR 195B enhancer region or a clone of the same region from the SU-DHL-5 cell line bearing a partial tandem duplication (“195B_MYC_Dup”) as indicated in (A) [two-tailed Student t test of biological replicates (n = 3); error bars are 95% CI]. **, P < 0.01; ***, P < 0.001; ns, not significant; P ≥ 0.05. FC, fold change; neg ctrl, negative control.

    Article Snippet: The following primary antibodies were used: MEF2B (Atlas Antibodies #HPA004734, RRID: AB_10963939), EBF (Santa Cruz Biotechnology #sc-137065, RRID: AB_2246405), E2A (Santa Cruz Biotechnology #sc-349, RRID: AB_675504), Bob1 (Santa Cruz Biotechnology #sc-955, RRID: AB_2166917), Oct-2 (Santa Cruz Biotechnology #sc-233, RRID: AB_2167205), Bcl-6 (Santa Cruz Biotechnology #sc-7388, RRID: AB_2063455), c-Myc (Cell Signaling Technology #5605, RRID: AB_1903938), CTCF (Cell Signaling Technology #3418, RRID: AB_2086791), c-Rel (Cell Signaling Technology #12707, RRID: AB_2721030), phosphoSer536-RelA (Cell Signaling Technology #3033, RRID: AB_331284), RelA (Cell Signaling Technology #8242, RRID: AB_10859369), NF-κB1/p50 (Cell Signaling Technology #12540, RRID: AB_2687614), NF-κB2/p52 (Cell Signaling Technology #3017, RRID: AB_10697356), and TBP (Abcam #ab818, RRID: AB_306337).

    Techniques: ChIP-sequencing, Binding Assay, Quantitative Proteomics, Quantitative RT-PCR, Expressing, Transduction, Clone Assay, Plasmid Preparation, Comparison, Two Tailed Test, Construct, Selection, Negative Control

    Fig. 7 | Stabilization of MEF2B C-terminal isoform-switch mutants by dis- sociation from the CUL3/KLHL12 complex. a Detection of MEF2B/CUL3/KLHL12 interaction by Flag (MEF2B) immunoprecipitation (IP) using nuclear extracts of SUDHL10 cells expressing Flag-HA-MEF2B-WT-isoA, -isoB or isoform-switch mutants. Lamin B1 was used as loading control. Quantifications were normalized to the immunoprecipitated MEF2B (HA). Fold changes were calculated relative to the MEF2B WT-isoA sample. b MEF2B IP followed by immunoblotting (IB) using nuclear extracts of isogenic OCI-Ly7 clones including controls (Neutral) and cells expres- sing MEF2B isoform-switch (P256fs and P273fs) mutants. IgG IP was used as nega- tive control. Quantifications were normalized to the immunoprecipitated MEF2B. Fold changes were calculated relative to the neutral clones. Samples are the same as in Fig. 5e. c Ubiquitination assay upon Flag (MEF2B) IP followed by IB using total extracts of HEK293T cells co-transfected with expression vectors for Flag-HA- MEF2B-WT-isoA, -isoB or isoform-switch mutants (P243fs, L269fs, and P273fs), and

    Journal: Nature communications

    Article Title: MEF2B C-terminal mutations enhance transcriptional activity and stability to drive B cell lymphomagenesis.

    doi: 10.1038/s41467-024-51644-8

    Figure Lengend Snippet: Fig. 7 | Stabilization of MEF2B C-terminal isoform-switch mutants by dis- sociation from the CUL3/KLHL12 complex. a Detection of MEF2B/CUL3/KLHL12 interaction by Flag (MEF2B) immunoprecipitation (IP) using nuclear extracts of SUDHL10 cells expressing Flag-HA-MEF2B-WT-isoA, -isoB or isoform-switch mutants. Lamin B1 was used as loading control. Quantifications were normalized to the immunoprecipitated MEF2B (HA). Fold changes were calculated relative to the MEF2B WT-isoA sample. b MEF2B IP followed by immunoblotting (IB) using nuclear extracts of isogenic OCI-Ly7 clones including controls (Neutral) and cells expres- sing MEF2B isoform-switch (P256fs and P273fs) mutants. IgG IP was used as nega- tive control. Quantifications were normalized to the immunoprecipitated MEF2B. Fold changes were calculated relative to the neutral clones. Samples are the same as in Fig. 5e. c Ubiquitination assay upon Flag (MEF2B) IP followed by IB using total extracts of HEK293T cells co-transfected with expression vectors for Flag-HA- MEF2B-WT-isoA, -isoB or isoform-switch mutants (P243fs, L269fs, and P273fs), and

    Article Snippet: The membranes were incubated overnight at 4 °C with the primary antibodies against MEF2B (custom23 and NBP303920, Novus Biologicals; 1:1000 dilution), pMEF2BS310 (custom; 1:1000 dilution), pMEF2BS324 (custom; 1:1000 dilution), Actin (A5441, Sigma-Aldrich; 1:3000 dilution), Vinculin (SAB4200080, SigmaAldrich; 1:2000 dilution), Lamin B1 (12586, Cell Signaling Technology; 1:1000dilution),HA (3724, Cell SignalingTechnology; 1:1000dilution), Myc (sc-40, Santa Cruz; 1:1000 dilution), Flag (P2983, Sigma-Aldrich; 1:1000 dilution), EGFP (2956, Cell Signaling Technology; 1:1000 dilution), Ubiquitinylated proteins (04-263, Sigma-Aldrich; 1:1000 dilution), K48-linkage Specific Polyubiquitin (8081, Cell Signaling Technology; 1:1000 dilution), K63-linkage Specific Polyubiquitin (5621, Cell Signaling Technology; 1:1000 dilution), V5 (13202, Cell Signaling Technology; 1:1000 dilution), SMARCA4 (49360, Cell Signaling Technology; 1:1000 dilution), CUL3 (A301-109A, Bethyl Laboratories; 1:1000 dilution) and KLHL12 (9406, Cell Signaling Technology; 1:1000 dilution).

    Techniques: Immunoprecipitation, Expressing, Control, Western Blot, Clone Assay, Ubiquitin Proteomics, Transfection

    Journal: Cell Reports Medicine

    Article Title: Targeting DNMT3A-mediated oxidative phosphorylation to overcome ibrutinib resistance in mantle cell lymphoma

    doi: 10.1016/j.xcrm.2024.101484

    Figure Lengend Snippet:

    Article Snippet: Rabbit polyclonal anti-MEF2B , ThermoFisher Scientific , PA5101143; RRID: AB_2850587.

    Techniques: Bacteria, Virus, Recombinant, Bicinchoninic Acid Protein Assay, RNA Extraction, Adhesive, Protease Inhibitor, Reporter Gene Assay, Mutagenesis, Luciferase, Purification, Sample Prep, Plasmid Preparation, Software