Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: TAL1 is lowly expressed in NSCLC. A DEGs screened out in the GSE18842 , GSE19188 , and GSE159857 datasets. B The intersection genes of TF and three GEO datasets were taken by Jvenn. C String database explored the interactions and took the top three scoring genes using the BottleNeck algorithm. D The UALCAN database found lowly expressed TAL1 in two subtypes of NSCLC. E RT-qPCR detected TAL1 mRNA expression in NSCLC cell lines (H1299, HCC827, A549, H460, and PC-9) and BEAS-2B. F WB detected TAL1 protein expression in NSCLC cell lines and BEAS-2B. Single-factor comparisons among multiple groups were performed using one-way ANOVA. **** p < 0.0001

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Quantitative RT-PCR, Expressing

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: Upregulation of TAL1 inhibits NSCLC progression. A RT-qPCR detected TAL1 expression in A549 and H460 cells after infection of OE-NC or OE-TAL1. B WB detected TAL1 protein levels in A549 and H460 cells after infection with OE-NC or OE-TAL1. C MTT analyzed the proliferative ability of A549 and H460 cells. D EdU assay validated the proliferation ability of A549 and H460 cells. E WB detected the expression of proliferation-related protein Ki67 and EMT-related proteins (E-cadherin, N-cadherin). F Wound healing assay detected the migration ability. G Transwell assay detected the invasion ability. H TUNEL detected the apoptosis rate. Comparisons were made among multiple groups using an unpaired t-test or a two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Quantitative RT-PCR, Expressing, Infection, EdU Assay, Wound Healing Assay, Migration, Transwell Assay, TUNEL Assay

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: TAL1 transcriptionally represses the expression of PKM2. A DEGs from three datasets, GSE18842 , GSE19188 , and GSE159857 , were taken. B KOBSA website for KEGG enrichment analysis. C String database analyzed the interoperable analysis of 81 genes in the pathway. D The top three gene clusters were obtained using the BottleNeck algorithm, the EcCentricity algorithm, and the Betweenness algorithm. E GEPIA database explored the correlation between PKM2 and TAL1. F The expression of PKM2 in LUAD and LUSC was predicted in the UALCAN database. G RT-qPCR analysis verified that PKM2 was highly expressed in NSCLC cell lines. H JASPAR predicted the binding site of TAL1 on the PKM2 promoter. I RT-qPCR assay for PKM2 expression in A549 and H460 cells. J Dual-luciferase assay detected the luciferase activity of PKM2. K . ChIP assay verified the enrichment of TAL1 at the PKM2 promoter. Comparisons were made using unpaired t-test, one-way, or two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Luciferase, Activity Assay

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: TAL1 inhibits NSCLC progression by blocking the glycolytic pathway through inhibition of PKM2. A Colony formation assay detected the colony-forming of A549 and H60 cells after 14 days of culture. B MTT analyzed the proliferation ability. C EdU assay verified the proliferation ability. D Wound healing assay detected migration. E Transwell assay detected the invasion ability. F TUNEL assay detected the apoptosis rate. G Cellular metabolism assay detected changes in lactate levels. H WB assay detected the protein levels of glycolysis-related proteins GLUT1, LDHA, and PDK1. Comparisons were made using unpaired t-tests or two-way ANOVA. ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Blocking Assay, Inhibition, Colony Assay, EdU Assay, Wound Healing Assay, Migration, Transwell Assay, TUNEL Assay

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: TAL1 blocks the glycolytic pathway by inhibiting PKM2 to slow NSCLC progression. A Tumor images and tumor weight of mice. B The tumor volume of mice was measured periodically. C WB detected the expression of TAL1 and PKM2 in the tumor tissues of mice. D IHC detected the expression of glycolysis-related proteins (GLUT1, LDHA, and PDK1), Ki67, and Cleaved-Caspase3 in the tumors of mice. E TUNEL detected apoptosis in tumor tissues of mice. Animal experiments had five mice in each group. Comparisons were made among multiple groups using one-way or two-way ANOVA. ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Expressing, TUNEL Assay

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: DNMT3B represses TAL1 expression by methylation modification. A UALCAN database to study the methylation level of TAL1 in LUAD and LUSC datasets. B Prediction of CpG islands in the promoter region of TAL1 by the MethPrimer database. C The correlation between TAL1 methylation levels and gene expression levels in LUAD or LUSC was predicted in the SMART database. D Correlation between DNMT3B expression and TAL1 methylation in LUAD and LUSC by Pearson’s analysis. E UALCAN database to study DNMT3B expression in LUAD and LUSC. F RT-qPCR analysis verified DNMT3B expression in NSCLC cells and BEAS-2B cells. G RT-qPCR verified the knockdown efficiency of DNMT3B. H RT-qPCR detected TAL1 expression in A549 and H460 cells. I Dual luciferase assay to detect TAL1 luciferase activity in A549 and H460 cells. J ChIP assay to validate DNMT3B enrichment at the promoter of TAL1 in A549 and H460 cells. K. qMSP assay to detect the methylation status of TAL1 in A549 and H460 cells. Comparisons were made using unpaired t-test, one-way, or two-way ANOVA. ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Expressing, Methylation, Modification, Gene Expression, Quantitative RT-PCR, Knockdown, Luciferase, Activity Assay

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: DNMT3B activates the glycolytic pathway by inhibiting TAL1 expression. A Colony formation assay to detect the proliferation of A549 and H460 cells after 14 days of culture. B EdU assay verified the proliferative ability of A549 and H460 cells. C , D The Transwell assay detected the migratory and invasive ability of A549 and H460 cells. E TUNEL assay detected apoptotic rate. F Cell metabolism assay detected changes in lactate levels. G WB detection of protein levels of glycolysis-related proteins GLUT1, LDHA, and PDK1. Comparisons were made among multiple groups using one-way or two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: Expressing, Colony Assay, EdU Assay, Transwell Assay, TUNEL Assay

Journal: Cell Division

Article Title: DNMT3B blocks TAL1-mediated PKM2 transcriptional repression to promote non-small cell lung cancer progression through inducing glycolysis

doi: 10.1186/s13008-025-00168-8

Figure Lengend Snippet: DNMT3B affects NSCLC progression in vivo by regulating the TAL1/PKM2 axis. A Tumor images and tumor weight of mice. B The tumor volume of mice was measured periodically. C WB detection of DNMT3B, TAL1, and PKM2 protein expression in tumor tissues of mice. D IHC detection of glycolysis-related proteins (GLUT1, LDHA, and PDK1), Ki67, and Cleaved-Caspase3 in the tumors of mice. E TUNEL detection of apoptosis in tumor tissues. Animal experiments had five mice in each group. One-way or two-way ANOVA was used for single-factor comparisons among multiple groups. ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: The used primary antibodies encompassed TAL1 (1:1000, A12927, ABclonal, Wuhan, Hubei, China), PKM2 (1:1000, 4053, CellSignaling, Beverly, MA, USA), DNMT3B (1:1000, 67259, CellSignaling), Ki67 (1:1000, ab92742, Abcam), E-cadherin (1:1000, A24874, ABclonal), N-cadherin (1:5000, ab76011, Abcam), glucose transporter type 1 (GLUT1, 1:1000, A6982, ABclonal), lactate dehydrogenase A (LDHA, 1:1000, 3582, CellSignaling), and pyruvate dehydrogenase kinase 1 (PDK1, 1:1000, NBP2-44132, Novus Biologicals, Littleton, CO, USA).

Techniques: In Vivo, Expressing, TUNEL Assay

Journal: Lab on a chip

Article Title: Multiplexed analysis of neural cytokine signaling by a novel neural cell-cell interaction microchip.

doi: 10.1039/d0lc00401d

Figure Lengend Snippet: Fig. 3 Immunocytology of neural rosettes formed on the NCCIM. (a) EB-derived rosettes in the dish and in the NCCIM. Cells are stained with DAPI, tight junction protein ZO1, NSC multipotency marker Nestin, neural ectoderm (Pax6), proliferation marker ki67, and neural progenitor (Sox1, Sox2) biomarkers as indicated. (b) Monolayer rosettes in the dish are dissociated and seeded into the NCCIM and form uniformly sized NSC rosette neurospheres by day 2. Cells are evaluated with similar biomarkers as (a). (c) Phase and fluorescence microscopy images of EB-derived rosettes in the PDMS microchambers of the NCCIM. Spontaneously formed rosettes in microwells results in three main size distribution categories: one large rosette, 3–4 medium size rosettes, and >5 small size rosettes. Cells are stained with ZO-1 (green) and Sox1 (red) antibodies. Scale bar is 50 μm.

Article Snippet: The antibodies, including Sox1 (R&D systems AF3360), Sox2 (R&D systems MAB2018), ZO-1 (ThermoFisher Scientific 33-9100), Ki67 (Abcam 15580), Nestin (R&D systems MAB1259), Pax6 (DSHB PAX6, Pax6 was deposited to the DSHB by Kawakami, A.) were added (1 : 1000) and incubated at 4 °C overnight.

Techniques: Derivative Assay, Staining, Marker, Fluorescence, Microscopy

Journal: Lab on a chip

Article Title: Multiplexed analysis of neural cytokine signaling by a novel neural cell-cell interaction microchip.

doi: 10.1039/d0lc00401d

Figure Lengend Snippet: Fig. 5 Viability and multipotency of rosettes on the NCCIM. (a) Flow diagram from seeding of cells into the PFMA that is combined with the MIST array to generate the assembled NCCIM. Cells were maintained within the NCCIM for 8 hours then evaluated by (b) propidium iodide (PI) staining of EB-derived rosettes and rosette neurospheres to visualize dead cells at hour 0 and hour 8. In (c) neural progenitor marker SOX1 is shown with adherens junction protein ZO-1 that highlights the center of polarized rosette cells. Cells maintain their multipotent state after NCCIM incubation for 8 hours. Scale bar is 50 μm.

Article Snippet: The antibodies, including Sox1 (R&D systems AF3360), Sox2 (R&D systems MAB2018), ZO-1 (ThermoFisher Scientific 33-9100), Ki67 (Abcam 15580), Nestin (R&D systems MAB1259), Pax6 (DSHB PAX6, Pax6 was deposited to the DSHB by Kawakami, A.) were added (1 : 1000) and incubated at 4 °C overnight.

Techniques: Staining, Derivative Assay, Marker, Incubation