anti lef1  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: A FZD4/LRP5 agonist restores pericyte coverage and vascular integrity by increasing PDGFB signaling

doi: 10.64898/2026.03.13.711629

Figure Lengend Snippet: (A) Quantification of Pdgfb from total retina RNA was normalized to beta-actin , n=9-11 retinas from 9-11 mice per group. Average +/− SE is shown. *P < 0.05 by 1-way ANOVA with Tukey’s post hoc test. (B) Quantification of Pdgfb from total retina RNA was normalized to gapdh , n=5-7 retinas from 5-7 mice per group. Average +/− SE is shown. *P < 0.05 by 1-way ANOVA with Tukey’s post hoc test. ( C ) Images of anti-LEF1 stain of parental bEnd.3 cells and a stable population of bEnd.3 cells overexpressing LEF1, generated by lentiviral transduction and selection with puromycin. Scale bar 100 µm. ( D ) Quantification of Pdgfb from total RNA of bEnd.3 cells after treatment with vehicle or F4L5.13. 2 technical replicates per biological replicate were averaged, n=3 biological replicates. Average +/− SE is shown. *P < 0.05 by unpaired Student’s t-test. ( E ) Quantification of Pdgfb from total RNA of stable bEnd.3-LEF1 cells after treatment with vehicle or F4L5.13. 2 technical replicates per biological replicate were averaged, n=3 biological replicates. Average +/− SE is shown. *P < 0.05 by unpaired Student’s t-test.

Article Snippet: LEF1 was stained in PFA-fixed cells using rabbit anti-LEF1 (Cell Signaling, #2230S 1:100) and goat anti-rabbit Alexa Fluor-555 (Invitrogen, #A21428).

Techniques: Staining, Generated, Transduction, Selection

Journal: Cell Death & Disease

Article Title: β-catenin mutation reprograms ketone body metabolism to drive hepatocellular carcinoma metastasis and resistance to ketogenic therapy via transcriptional activation of OXCT1

doi: 10.1038/s41419-026-08457-y

Figure Lengend Snippet: A Correlation between OXCT1 and CTNNB1 (encoding β-catenin) RNA expression in HCC samples from the TCGA dataset. B Western blot analyzed the protein levels of β-catenin and OXCT1 in HCCLM3 and Huh7 cells. C , D The mRNA and protein levels of β-catenin and OXCT1 were analyzed by Western blot or RT-qPCR in HCCLM3-β-catenin S33Y (β-catenin) or control (CON) cells. E , F The mRNA and protein levels of β-catenin and OXCT1 were analyzed by Western blot or RT-qPCR in Huh7 cells transfected with shRNA of CTNNB1 (shCTNNB) or no-targeting control (shCON). G Correlation between OXCT1 and LEF1 mRNA expression in HCC samples from the TCGA database. H Western blot analyzed protein levels of OXCT1 and LEF1 in HCCLM3 cells transfected with LEF1 plasmid or empty vector (EV). ( I ) Western blot analyzed protein levels of OXCT1 in HCCLM3 cells transfected with LEF1 shRNA (shLEF1) or β-catenin S33Y vector. J Co-IP assay demonstrating the interaction between β-catenin and LEF1 in Huh7 cells. K ChIP assay was performed with LEF1 antibody. IgG was used as the negative control. Chip-PCR was conducted at the promoter regions of OXCT1 with Huh7 cells. L Dual-luciferase reporter assays in HEK293 cells. Cells were co-transfected with a wild-type (PGL3-OXCT1-WT) or mutant (PGL3-OXCT1-MT) OXCT1 promoter reporter, along with expression plasmids for β-catenin, LEF1, TCF4+β-catenin, or LEF1 shRNA, as indicated. Luciferase activity was measured 48 h post-transfection. M Schematic of the OXCT1 promoter region. The consensus LEF1-binding site (5′-ACTTTGAACT-3′) is indicated. Data are presented as mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001, ns, not significant.

Article Snippet: Primary antibodies against the following proteins were used: β-actin (#20536-1-AP, Proteintech, China), OXCT1 (#12175-1-AP, Proteintech, China), β-catenin (#8480, Cell Signaling Technology, USA), PPARα (#66826-1-Ig, Proteintech, China), HMGCS2 (#ab137043, Abcam, USA), BDH1 (#15417-1-AP, Proteintech, China), LEF1 (#14972-1-AP, Proteintech, China), MMP2 (#10373-2-AP, Proteintech, China), Snai1 (13099-1-AP, Proteintech, China), Vimentin(#60330-1-Ig, Proteintech, China), E-cadherin (HY- P81271 , MCE, China), STAT3 (#9139, Cell Signaling Technology, USA), p-STAT3 (#9145, Cell Signaling Technology, USA).

Techniques: RNA Expression, Western Blot, Quantitative RT-PCR, Control, Transfection, shRNA, Expressing, Plasmid Preparation, Co-Immunoprecipitation Assay, Negative Control, Luciferase, Mutagenesis, Activity Assay, Binding Assay