Journal: BMC Biotechnology

Article Title: A TXNIP-driven bioluminescent reporter for high-throughput discovery of glycolytic inhibitors against renal cell carcinoma

doi: 10.1186/s12896-026-01111-7

Figure Lengend Snippet: 2-DG upregulates MLXIP and TXNIP expression in A498 cells. ( A ) Volcano plot of differentially expressed genes (DEGs). ( B ) Top 10 upregulated genes from RNA-seq analysis. ( C ) TXNIP mRNA detected by RT-qPCR in A498 cells after 48 h of treatment with 2-DG (0, 5, 10, and 15 mM). ( D ) Protein expression of TXNIP and MLXIP was detected by Western blotting in A498 cells treated with 2-DG at 0, 5, 10, and 15 mM for 48 h. The 0 mM group served as the vehicle control and contained an equal volume of DMSO. ( E ) MLXIP and TXNIP mRNA expression analyzed by RT-qPCR in A498 cells 48 h after transfection with MLXIP plasmid (0, 2, and 4 µg), where 0 µg MLXIP plasmid corresponds to the empty vector plasmid used as control. Data are mean ± SEM ( n = 3); Statistical significance was analyzed by one-way ANOVA with Dunnett’s multiple comparisons; **** p < 0.0001, *** p < 0.001

Article Snippet: The primary antibodies used included: rabbit polyclonal anti-MLXIP antibody (Cat. No. 13614-1-AP, RRID: AB_2282153, Proteintech, Wuhan, China), rabbit monoclonal anti-TXNIP antibody (Cat. No. A11682, RRID: AB_2935845, Nature Biosciences, Hangzhou, China), and rabbit monoclonal anti-Vinculin antibody (Cat. No. A2752, RRID: AB_2863020, Abclonal, Wuhan, China).

Techniques: Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Control, Transfection, Plasmid Preparation

Journal: BMC Biotechnology

Article Title: A TXNIP-driven bioluminescent reporter for high-throughput discovery of glycolytic inhibitors against renal cell carcinoma

doi: 10.1186/s12896-026-01111-7

Figure Lengend Snippet: Characterization of a stable RCC reporter cell line with TXNIP promoter-driven luciferase expression. ( A ) Schematic diagrams of pGL4.19-TXNIP-Pro-Luc2 constructs. The TXNIP promoter fragment, spanning from − 1166 bp to + 312 bp relative to the transcription start site (TSS), was cloned into the pGL4.19-Luc2 vector to drive luciferase expression. ( B ) A498-TXNIP-Pro-Luc2 cells or A498-Luc2 cells were lysed for luciferase activity analysis. ( C ) A498-TXNIP-Pro-Luc2 and A498-Luc2 cells were imaged using the IVIS Lumina LT system to obtain flux measurements (left panel, images). Quantified flux data were averaged ( n = 3) and plotted (right panel, graph). ( D , E ) After transfection of the MLXIP plasmids into A498-TXNIP-Pro-Luc2 ( D ) or A498-Luc2 cells ( E ) for 48 h, imaging was performed (left panel, images). Quantified flux data were averaged ( n = 3) and plotted (right panel, graph). ( F ) After transfection of the MLXIP plasmids into A498-TXNIP-Pro-Luc2 or A498-Luc2 cells for 48 h, the cells were lysed for luciferase activity analysis. ( G , H ) A498-TXNIP-Pro-Luc2 cells ( G ) and A498-Luc2 cells ( H ) were serially diluted, placed into wells of a 96-well plate, and immediately imaged. Quantified flux data were averaged ( n = 3) and plotted. Data are mean ± SEM ( n = 3); Statistical significance for B and C was analyzed by one-way ANOVA; for D , E , and F , it was analyzed by one-way ANOVA with Dunnett’s multiple comparisons test; **** p < 0.0001

Article Snippet: The primary antibodies used included: rabbit polyclonal anti-MLXIP antibody (Cat. No. 13614-1-AP, RRID: AB_2282153, Proteintech, Wuhan, China), rabbit monoclonal anti-TXNIP antibody (Cat. No. A11682, RRID: AB_2935845, Nature Biosciences, Hangzhou, China), and rabbit monoclonal anti-Vinculin antibody (Cat. No. A2752, RRID: AB_2863020, Abclonal, Wuhan, China).

Techniques: Luciferase, Expressing, Construct, Clone Assay, Plasmid Preparation, Activity Assay, Transfection, Imaging