In HUVEC on Geltrex, 10 ng/ml IMD-0354 for 24 h reduced junction formation, tubule number and total tubule length. Propidium-iodide staining showed no significant increase in cell death in this assay. [1]
IMD-0354 reduced radiolabeled glutamine uptake in A431 carcinoma cells and A375 melanoma cells. This cellular transport result does not establish direct transporter binding or selectivity. [2]
NF-κB localization and phospho-IκBα in HUVEC
Cell-based study · Serum-starved human umbilical vein endothelial cells (HUVEC) [1]
Compared with DMSO vehicle, IMD0354-treated cells showed less nuclear p65 localization and less phospho-IκBα staining. These are qualitative cellular pathway observations, not isolated-enzyme inhibition or direct-binding measurements.
Literature-Reported Treatment Conditions
- Treatment concentration
- 10 ng/ml
- Exposure duration
- 24 h treatment; Figure 1 describes 24 h serum-starved HUVEC
- Method
- Immunofluorescent localization of NF-κB p65 and phospho-IκBα; DAPI nuclear stain
- Vehicle
- DMSO; Figure 1 vehicle control 1 µl/ml
- Reported limits
- Figure 1 does not state a replicate count or quantitative effect size. HUVEC-specific p65/phospho-IκBα staining steps are not separately detailed; shared immunohistochemistry methods were read.
Methods and source locations
Compound reported in the paper: IMD-0354; Compound label in the paper: IMD0354
Compound-level experimental evidence only; no salt conversion, clinical efficacy or current-lot validation.
Results Sec3; Figure 1a-b and caption; Methods Sec16 and Sec28
Nuclear NF-κB p65 localization and phospho-IκBα staining: Immunofluorescence; representative images, no numeric effect inferred — Figure 1a-b; Results Sec3
HUVEC tube formation with concurrent cell-death assessment
Cell-based study · HUVEC on Geltrex [1]
IMD0354 reduced tube-network junctions, tubule number and total tubule length compared with DMSO. The concurrent propidium-iodide endpoint showed no significant increase in cell death under this assay condition.
Literature-Reported Treatment Conditions
- Treatment concentration
- 10 ng/ml
- Exposure duration
- 24 h
- Method
- Geltrex tube-formation imaging and AngioSys quantification; calcein-AM and propidium iodide staining
- Vehicle
- DMSO; Figure 2 control 1 µl/ml
- Reported limits
- Caption gives n = 8, without establishing eight independent donors or biological experiments. Exact effect sizes were not tabulated; none inferred from bars.
Methods and source locations
Compound reported in the paper: IMD-0354; Compound label in the paper: IMD0354
Compound-level experimental evidence only; no salt conversion, clinical efficacy or current-lot validation.
Results Sec4; Figure 2c-g and caption; Methods Sec16, Sec18, Sec31
Junction number, tubule number and total tubule length: Image analysis; Student t test as described for these comparisons; values described as mean ± SD — Figure 2c-f and caption; Results Sec4; Methods Sec18 and Sec31
Propidium-iodide-positive cell count: Concurrent live/dead fluorescence imaging; this endpoint does not establish general safety — Figure 2c,g and caption; Results Sec4
Cellular inhibition of glutamine uptake
Cell-based study · A431 human epidermoid carcinoma cells and A375 human melanoma cells [2]
Radiotracer uptake was reduced in A431 cells at 1 h and 24 h, and concentration-dependent inhibition was observed in A431 and A375 cells in the 1 h comparison. This is a cellular transport endpoint; it does not prove direct SLC1A5 binding or transporter selectivity.
Literature-Reported Treatment Conditions
- Treatment concentration
- Figure 3a: 5 or 10 µM in A431; Figure 3b: 0.63, 2.5 or 10 µM in A431 and A375
- Exposure duration
- Figure-specific treatment times: 1 h or 24 h in 3a; 1 h in 3b
- Method
- 3H-glutamine uptake quantified by scintillation counting
- Vehicle
- DMSO control (Figure 3); concentration not specified for this experiment in the read main text
- Reported limits
- Methods describe a generic 15 min compound preincubation and 15 min tracer uptake; figure-specific pretreatments are retained without reconstructing a unified schedule. No IC50 is inferred from the plotted doses. Figure caption n = 3; main statistics states cell culture experiments performed three times.
Methods and source locations
Compound reported in the paper: IMD-0354; Compound label in the paper: IMD-0354
Compound-level experimental evidence only; no salt conversion, clinical efficacy or current-lot validation.
Figure 3a-b and caption; Results: IMD-0354 inhibits glutamine uptake; Methods: 3H-Glutamine uptake assay and Reagents
Relative 3H-glutamine uptake: Scintillation-based uptake assay; no exact percentages estimated from bar heights — Figure 3a-b and caption; Results glutamine-uptake subsection; Methods 3H-Glutamine uptake assay
