Journal: bioRxiv

Article Title: ATGL-catalyzed lipid catabolism promotes DNA repair

doi: 10.64898/2026.04.03.716381

Figure Lengend Snippet: A) AML12 cells transduced with ATGL or GFP control adenoviruses were treated with 30 μM etoposide for the indicated times and assayed for γH2Ax. B) Quantification of A. *p<0.05, **p<0.01. C) AML12 cells transduced with ATGL or GFP control adenoviruses were irradiated (10 Gy) and harvested at the indicated times post-IR. D) Quantification of C. **p<0.01. E) Primary MEFs were isolated from WT or ATGL knock-in (AKI) mice, followed by etoposide incubation for 6 hours. Immunofluorescence was performed for γH2Ax, and cells were imaged using the Agilent Gen5 Cytation. Representative γH2Ax images and foci distribution are shown. F) Cells were treated with DGAT inhibitors for 24 hours, followed by ATGL adenoviral transduction, followed by 24 hours of etoposide treatment. Western blotting for γH2Ax, β-actin, and ATGL. G) AML12 cells were treated as in F. The Comet assay was performed on cells after 24 hours of etoposide. Comets were quantified with OpenComet software. A minimum of 180 comets were quantified per condition. Statistics: one-way ANOVA with Tukey’s post hoc test. ****p<0.0001. H) γH2Ax results and quantification from tissues harvested from E2A Cre-ATGL (AKI) mice irradiated with 7.5 Gy + 4-hour recovery. Mice were of mixed sexes, n=4 mice. P-values from one-way ANOVA are displayed

Article Snippet: E2A Cre mice (B6.FVB-Tg(EIIa-Cre)C5379Lmgd/J, Jackson Labs) were of B6.FBV background.

Techniques: Transduction, Control, Irradiation, Isolation, Knock-In, Incubation, Immunofluorescence, Western Blot, Single Cell Gel Electrophoresis, Software

Journal: Redox Biology

Article Title: LDB1 represses fetal hemoglobin expression by enhancing BCL11A transcription

doi: 10.1016/j.redox.2026.104070

Figure Lengend Snippet: The LDB1 complex containing LMO2 and GATA1 directly enhances the transcription of the Bcl11a gene. ( A-E ) ChIP-qPCR assays were performed to assess the occupancy of putative LDB1 complex component proteins, including LDB1, LMO2, GATA1, TAL1, and E2A, at the Bcl11a locus in E14.5 WT and KO FLCs. n = 3. Control, isotype control Ab; LDB1, anti-LDB1 Ab; LMO2, anti-LMO2 Ab; GATA1, anti-GATA1 Ab; TAL1, anti-TAL1 Ab, E2A, anti-E2A Ab; WT, E14.5 FLCs; KO, E14.5 KO FLCs. Statistical significance was assessed by one-way ANOVA with Tukey HSD analysis. Mean values not sharing the same superscript letter ( a, b ) differ significantly at P < 0.05. ( F and G ) Dual-luciferase reporter assays were performed using the 210 bp region covering primer #3 ( Bcl11a intron #3) or the 225 bp region covering primer #4 ( Bcl11a intron #4) at Bcl11a locus. The relative luciferase activity of each construct containing either Bcl11a intron #3 or Bcl11a intron #4 was measured in HEK293T cells transfected with the indicated combination of each expression vector encoding Ldb1 , Lmo2 , or Gata1 cDNA. The schematic diagrams of each cloned enhancer region ( F ) and the results of the luciferase reporter assays ( G ) are presented. Ldb1 , expression vector encoding Ldb1 cDNA; Lmo2 , expression vector encoding Lmo2 cDNA; Gata1 , expression vector encoding Gata1 cDNA; ◯, transfected cells; ☓, untransfected cells. n = 5. Statistical significance was assessed by one-way ANOVA with Tukey HSD analysis. Mean values not sharing the same superscript letter ( a, b, c ) differ significantly at P < 0.05. ( H and I ) ChIP-qPCR assays were performed to assess LDB1 occupancy at the human BCL11A locus in K562 cells. The locations of each indicated primer set within the BCL11A locus are shown in the schematic diagram ( H ) along with the corresponding ChIP-qPCR results ( I ). n = 3. Mock K562, empty vector transfected K562 cells; KO K562, human LDB1 knockout K562 cells; Control, isotype control Ab; LDB1, anti-LDB1 Ab. Statistical significance was assessed by two-tailed Student's t -test. ∗ P < 0.05. All data are presented as the mean ± SEM.

Article Snippet: Sheared chromatin was immunoprecipitated using 2 μg of the following Abs per sample: anti-LDB1 (Santa Cruz Biotechnology sc-365074) and its isotype control (R&D Systems MAB004), anti-GATA1 (Santa Cruz Biotechnology sc-265) and its isotype control (Santa Cruz Biotechnology, Inc. sc-3883), anti-TAL1 (Santa Cruz Biotechnology sc-393287), anti-LMO2 (Santa Cruz Biotechnology sc-65736), anti-E2A (Santa Cruz Biotechnology sc-416) and their isotype control (Thermo Fisher Scientific 14-4714-85).

Techniques: ChIP-qPCR, Control, Luciferase, Activity Assay, Construct, Transfection, Expressing, Plasmid Preparation, Clone Assay, Knock-Out, Two Tailed Test