Journal: The EMBO Journal

Article Title: TXNIP mediates LAT1/SLC7A5 endocytosis to limit amino acid uptake in cells entering quiescence

doi: 10.1038/s44318-025-00608-9

Figure Lengend Snippet: ( A ) WT, TXNIP KO and TXNIP KO cells reconstituted with TXNIP were harvested, permeabilized and fixed. The DNA content was analysed by PI staining to assess the cell cycle profile by FACS ( n = 3, N > 4000 cells, SD). ( B ) Increase in confluency of the indicated cells, after refeeding serum starved cells, was measured by continuous live cell microscopy using a Incucyte S3 system in an incubator for 48 h ( n = 6, SEM). ( C ) Competition assay of TXNIP KO + mCherry and TXNIP KO + GFP-TXNIP. 100 000 TXNIP KO + mCherry and 100 000 TXNIP KO + GFP-TXNIP were co-seeded and the cell number was measured by continuous live cell microscopy in an Incucyte incubator for 72 h. Scale bar = 400 µm ( n = 3, SEM). Quantification of TXNIP KO + mCherry and TXNIP KO + GFP-TXNIP cell number fold increase over time and quantification in % of total cells ( n = 3, two-way ANOVA, Sidak’s multiple comparisons test). ( D ) Analysis of cell death during quiescence of WT, TXNIP KO and SLC7A5 KO single knock out cells, and SLC7A5 KO TXNIP KO double knock out cells. 100,000 cells were seeded in growth medium ( + serum). After 24 h the medium was changed to starvation medium (-serum) containing propidium iodide (1 µg/ml). PI positive cells were measured by continuous live cell microscopy in an Incucyte incubator for 80 h. Scale bar = 400 µm. ( n = 3, SEM). ( E ) Model of TXNIP mediated downregulation of SLC7A5 during entry into quiescence. .

Article Snippet: IncuCyte S3 software , Sartorius , Version 2022B.

Techniques: Staining, Microscopy, Competitive Binding Assay, Knock-Out

Journal: The EMBO Journal

Article Title: TXNIP mediates LAT1/SLC7A5 endocytosis to limit amino acid uptake in cells entering quiescence

doi: 10.1038/s44318-025-00608-9

Figure Lengend Snippet: ( A ) The proliferation of the indicated cells was determined using CASY cell counting during the indicated time. The relative increase in cell number is normalized to 0 h and show as fold increase. Bar charts represent the mean value ( n > 3, SD). ( B ) Analysis of cell death during quiescence of WT, TXNIP KO TXNIP KO (+ DMSO), TXNIP KO reconstituted with GFP-TXNIP and TXNIP KO treated with JPH203 (10 µM). 100.000 cells were seeded in growth medium (+ serum). After 24 h the medium was changed to starvation medium (-serum) containing propidium iodide (1 µg/ml). At the same time JPH203 (10 µM) was added. Cells and PI positive cells were continuously monitored for 80 h using live cell microscopy in an Incucyte incubator. Scale bar = 400 µm ( n = 6, SEM).

Article Snippet: IncuCyte S3 software , Sartorius , Version 2022B.

Techniques: Cell Counting, Microscopy