e47  (Santa Cruz Biotechnology)

Journal: Bone Research

Article Title: PTH induced osteoblast Slit3 to decrease aberrant sensory innervation in degenerated vertebral endplates to relieve low back pain in mice

doi: 10.1038/s41413-025-00488-z

Figure Lengend Snippet: Transcriptional mechanism underpinning Slit3 secretion. a mRNA expression levels of Ets1, E47, FoxJ2 , and FoxA2 genes in MC3T3 cells cultured in osteoblast differentiation-inducing medium and treated with vehicle or PTH (100 nmol/L) for 3 days ( n = 3, t -test). b , c Protein expression level of E47, FoxA2 in MC3T3 cells cultured in stimulated medium and treated with PTH or Veh (PBS) for 3 days ( n = 3, t -test). Representative images depicting E47 IF staining (green) in lumbar vertebral body and endplate sections ( d ), quantitative analysis of the number of E47 + cells in the vertebral body ( e ) and endplate ( f ) of aged mice treated with Veh or PTH ( n = 5, t -tset). Scale bar: 100 µm. Representative images showing FoxA2 IF staining (red) in lumbar vertebral body and endplate sections ( g ), quantitative analysis of the number of FoxA2 + cells in the vertebral body ( h ) and endplate ( i ) of aged mice treated with PTH or vehicle for 2 months ( n = 5, t -tset). Scale bar: 100 µm. j Relative fold enrichment of the E47 binding site in the Slit3 promoter region from stimulated MC3T3 cells treated with vehicle or PTH (100 nmol/L) for 3 days ( n = 3, two-way ANOVA with Sidak’s multiple comparisons test). k Relative fold enrichment of the FoxA2 binding site in the Slit3 promoter region from stimulated MC3T3 cells treated with vehicle or PTH (100 nmol/L) for 3 days ( n = 3, two-way ANOVA with Sidak’s multiple comparisons test). * P < 0.05, ** P < 0.01, *** P < 0.005, **** P < 0.001

Article Snippet: Antibodies targeting E47 (sc-416, Santa Cruz), FoxA2 (22474-1-AP, Proteintech) were utilized.

Techniques: Expressing, Cell Culture, Staining, Binding Assay

Journal: eLife

Article Title: Degradation of LMO2 in T cell leukaemia results in collateral breakdown of transcription complex partners and causes LMO2-dependent apoptosis

doi: 10.7554/eLife.106699

Figure Lengend Snippet: ( A ) The T cell lines Jurkat (LMO2-), KOPT-K1 (LMO2+), and P12-Ichikawa (LMO2+) cells were infected with lentivirus packaging plasmids and transfer vector (TLCV2-VH576-L10-CRBN) for 16 hr followed by 2 μg/ml doxycycline induction for 9, 24, and 48 hr. Western blotting analysis was used to detect LMO2, endogenous CRBN, and VH576-L10-CRBN levels after lentivirus infection. α-Tubulin was used as an internal loading control for western blotting analysis. ( B ) The level of LMO2 and proteins associated with the LMO2 transcription complex (TAL1, E47, Lyl-1, GATA3, and LDB1) was determined after KOPT-K1 cells were infected with TLCV2-VH576-L10-CRBN followed by 2 μg/ml doxycycline induction for 9, 24, and 48 hr. Cyclophilin-b was used as an internal loading control. Figure 1—source data 1. Western blot data with label shows the effect of lentiviral expressed biodegrader to the LMO2 multi-protein complex in T cell lines. Figure 1—source data 2. Western blot raw data shows the effect of lentiviral expressed biodegrader to the LMO2 multi-protein complex in T cell lines. Figure 1—source data 3. Western blot data with label shows the level of LMO2 and proteins associaed with the LMO2 transcription complex. Figure 1—source data 4. Western blot raw data shows the level of LMO2 and proteins associaed with the LMO2 transcription complex.

Article Snippet: Antibody , Anti-E2A (E47) (Rabbit polyclonal) , Cell Signaling , Cat# 4865, RRID: AB_10560512 , WB (1:1000).

Techniques: Infection, Plasmid Preparation, Western Blot, Control

Journal: eLife

Article Title: Degradation of LMO2 in T cell leukaemia results in collateral breakdown of transcription complex partners and causes LMO2-dependent apoptosis

doi: 10.7554/eLife.106699

Figure Lengend Snippet: LMO2-expressing (KOPT-K1 and CCRF-CEM) and non-expressing T cells (Jurkat and DND-41) were treated with antibody-derived (Abd) degraders, and protein extracts were prepared for western blot, detecting LMO2, TAL1, E47, Lyl-1, GATA3, LDB1 proteins (cyclophilin-b was used as an internal loading control) after cells were treated with different concentrations of Abd degraders at 0, 5, 10, 15, and 20 μM for 24 hr. ( A ) Western blotting analysis with KOPT-K1 and CCRF-CEM extracts and ( B ) western blotting analysis with Jurkat and DND-41 extracts. ( C ) KOPT-K1 and CCRF-CEM were treated with Abd degraders at 20 μM for 2, 4, 6, and 24 hr. Western blotting analysis data show LMO2, TAL1, and E47 proteins expression in KOPT-K1 and CCRF-CEM that was affected by treatment. ( D ) The involvement of proteasome machinery in protein complex degradation was investigated in KOPT-K1 and CCRF-CEM with either proteasome inhibitors or by competing the potential of the Abd degraders with free E3 ligase ligand. Western blot data show LMO2, TAL1, and E47 expression in KOPT-K1 and CCRF-CEM after the treatment with or without inhibitors followed by Abd compounds. Inhibitors used were the proteasome inhibitor epoxomicin (0.8 μM), or CRBN inhibitors (10 μM thalidomide) or free VHL ligand. Cyclophilin-b was used as an internal loading control for western blotting analysis. Figure 3—source data 1. Western blot data with label shows ptoteins associated within the LMO2 transcription complex in KOPT-K1 and CCRF-CEM. Figure 3—source data 2. Western blot raw data shows ptoteins associated within the LMO2 transcription complex in KOPT-K1 and CCRF-CEM. Figure 3—source data 3. Western blot data with label shows ptoteins associated within the LMO2 transcription complex in Jurkat and DND-41. Figure 3—source data 4. Western blot raw data shows ptoteins associated within the LMO2 transcription complex in Jurkat and DND-41. Figure 3—source data 5. Western blot data with label shows ptoteins associated within the LMO2 transcription complex in KOPTK1- and CCRF-CEM after treated with Abd-degraders at 20 μM. Figure 3—source data 6. Western blot raw data shows ptoteins associated within the LMO2 transcription complex in KOPTK1- and CCRF-CEM after treated with Abd-degraders at 20 μM. Figure 3—source data 7. Western blot data with label shows the involvement of proteasome machinery in protein complex degradation. Figure 3—source data 8. Western blot raw data shows the involvement of proteasome machinery in protein complex degradation.

Article Snippet: Antibody , Anti-E2A (E47) (Rabbit polyclonal) , Cell Signaling , Cat# 4865, RRID: AB_10560512 , WB (1:1000).

Techniques: Expressing, Derivative Assay, Western Blot, Control

Journal: eLife

Article Title: Degradation of LMO2 in T cell leukaemia results in collateral breakdown of transcription complex partners and causes LMO2-dependent apoptosis

doi: 10.7554/eLife.106699

Figure Lengend Snippet: The crystal structure of the LMO2-TAL1-E47 complex, adapted from , is shown with LMO2 contacting the LID domain of LDB1, the TAL1 and E47 bHLH domains, and DNA. LMO2 is shown in grey; the VH576 interaction domain in pink ; LID in orange; TAL1 in blue, and E47 in green. Lysine residues in TAL1 and E47 are in yellow and salmon, respectively.

Article Snippet: Antibody , Anti-E2A (E47) (Rabbit polyclonal) , Cell Signaling , Cat# 4865, RRID: AB_10560512 , WB (1:1000).

Techniques:

Journal: eLife

Article Title: Degradation of LMO2 in T cell leukaemia results in collateral breakdown of transcription complex partners and causes LMO2-dependent apoptosis

doi: 10.7554/eLife.106699

Figure Lengend Snippet: Western blot data showed LMO2 and LMO2 protein complex (TAL1, E47, LYL1, LDB1, and GATA3) level after the treatment with 20 μg/ml cycloheximide from 0 to 24 hr ( A ). The level of protein remaining is presented as a relative to protein level at 0 hr ( B ). Data represent mean + SEM (n=3). Figure 3—figure supplement 1—source data 1. Western blot data with label shows half-lives of LMO2 and LMO2 protein complex in KOPT-K1. Figure 3—figure supplement 1—source data 2. Western blot raw data shows half-lives of LMO2 and LMO2 protein complex in KOPT-K1.

Article Snippet: Antibody , Anti-E2A (E47) (Rabbit polyclonal) , Cell Signaling , Cat# 4865, RRID: AB_10560512 , WB (1:1000).

Techniques: Western Blot

Journal: Nucleic acids research

Article Title: Integrated multi-omics analysis of PBX1 in mouse adult neural stem- and progenitor cells identifies a transcriptional module that functionally links PBX1 to TCF3/4.

doi: 10.1093/nar/gkae864

Figure Lengend Snippet: Figure 5. bHLH class I proteins are PBX1 interaction partners. ( A–C ) Similar peak distribution of neurode v elopmental ChIP-seq according to annotation with ChIPseeker: ( A ) PBX1 (V-SVZ aNS), ( B ) TCF3 (E12.5 NSC), ( C ) TCF4 (NS5 NSC). ( D ) ChIP-seq profiles of PBX1, TCF3 and TCF4 show similar pattern and distribution at example locus Itgb5 . Colored bars represent called peaks, grey arrow direction of transcription. ( E ) CentriMo analysis of PBX1 ChIP-seq peaks demonstrates PBX1 and bHLH class I motif co-occurrence at peak center. ( F, F’ ) Secondary binding analysis (SpaMo) of intersecting PBX1 (V-SVZ aNS) and TCF3 (E12.5 NSC) ChIP-seq peaks with MEME-ChIP; TALE-HD and bHLH class I motifs are spaced with a gap of 18 nt ( P = 8.23e-2; F), on both strands and in all directions ( F’ ). ( G ) PBX1 in aNS cells co-precipitates TCF3 / E47-FLAG in Co-IP of HEK293T protein lysate. ( H-H”’ ) P ro ximity ligation assa y f or PBX1 and TCF3 / E47-FLAG; ( H ) Script-based automated quantification re v ealing significant enrichment of punctate PLA signal in Tcf3 / E47-flag o v ere xpressing cells compared to empty vector control treatment, paired Student’s t -test ( P = 0.032); ( H’–H”’ ) Example staining showing speckled PLA signal in GFP + cells (arrow heads) as opposed to GFP low / - cells.

Article Snippet: Correlation analysis was erformed with the Pearson correlation test in R Studio Verion 1.3.1093. o-immunoprecipitation (co-IP) o assess interaction between TCF3 and PBX1, p3xFlagcf3 / E47 (murine Tcf3 splice variant E47 ; Addgene #34585) r p3xFlag-Tcf3 / E47aa290-648 (coding for amino acids 290– 48 of mouse E47) were introduced into HEK293T cells by alcium phosphate transfection.

Techniques: ChIP-sequencing, Binding Assay, Co-Immunoprecipitation Assay, Ligation, Plasmid Preparation, Control, Staining

Journal: Nucleic acids research

Article Title: Integrated multi-omics analysis of PBX1 in mouse adult neural stem- and progenitor cells identifies a transcriptional module that functionally links PBX1 to TCF3/4.

doi: 10.1093/nar/gkae864

Figure Lengend Snippet: Figure 6. PBX1-TCF interaction in neural progenitor cells. ( A ) Proportion of PSA-NCAM positive-neurons generated from aNS upon Pbx1 , Tcf3 and Tcf4 kd in aNS and quantified by FACS analysis ( n = 3, P < 0.05, paired t wo-t ailed Student’s t-test). ( B ) Venn diagram of commonly dysregulated genes upon Pbx1 , Tcf3 and Tcf4 kd identified by RNA-seq. ( C ) Heatmap and cluster analysis (euclidean clustering based on average) of commonly Pbx1 , Tcf3 and Tcf4 kd dysregulated genes in RNA-seq with log 2 FoldChange > 0.3. ( D ) Commonly downregulated genes upon Pbx1 and Tcf3 kd partake in replication-associated interaction network (STRING analysis). ( D ’–D ”’ ) Expression of Tcf 3 correlated with that of Brca2 , Rbl1 and Timeless in scRNA-seq of V-SVZ ( 75 ); Pearson Correlation test. ( E, E’ ) ChIP-qPCR of PBX1 ( E ) and TCF3 transcript variant E47-FLAG ( E’ ) at Olig2 upstream enhancer and promotor, Cited2 , Rbl1 , Prom1 , Myog and Dnm2 in Tcf3 / E47-Flag transduced aNS cells. Statistic analysis with paired Student’s t -test (FLAG IP: TCF3 / E47-FLAG vs. empty vector control, PBX1 IP: respective gene against neg. ctrl. Myog, n = 3–4, * P < 0.05).

Article Snippet: Correlation analysis was erformed with the Pearson correlation test in R Studio Verion 1.3.1093. o-immunoprecipitation (co-IP) o assess interaction between TCF3 and PBX1, p3xFlagcf3 / E47 (murine Tcf3 splice variant E47 ; Addgene #34585) r p3xFlag-Tcf3 / E47aa290-648 (coding for amino acids 290– 48 of mouse E47) were introduced into HEK293T cells by alcium phosphate transfection.

Techniques: Generated, RNA Sequencing, Expressing, ChIP-qPCR, Variant Assay, Plasmid Preparation, Control