RepSox (3 μM, 8 days) induced lipid accumulation, UCP1 expression and increased respiration in mouse embryonic fibroblasts. Rosiglitazone alone at 1 μM did not induce adipocytes under these conditions, while its combination with RepSox enhanced several differentiation readouts. [2]
In CCK-8 assays, the reported 96 h RepSox IC50 values were 140 μM in HOS cells and 149.3 μM in 143B cells. These values describe cellular viability endpoints and should remain distinct from receptor-level potency. [1]
RepSox responses depend on the cellular context. In primary AML/MDS cultures, 16 μM RepSox for 6 days reduced the mean decline in the CD34-positive fraction versus DMSO (33% ± 7% versus 54% ± 7%, mean ± SE; P1–P7; p < .005). The healthy-donor comparator did not show the same preservation. [3]
CRL-2097 dermal fibroblasts showed a concentration-dependent reversal: 100 nM RepSox lowered basal α-SMA and calponin expression, whereas higher micromolar concentrations up to 10 μM increased these markers after culture until day 4. Antifibrotic effects therefore require model- and concentration-specific interpretation. [4]
Brown adipocyte characteristics induced in mouse fibroblasts
Cell-based study · Primary C57BL/6 mouse embryonic fibroblasts (MEFs) [2]
RepSox alone induced lipid accumulation, UCP1 expression and higher basal, uncoupled and maximal respiration versus DMSO. Rosiglitazone alone was ineffective for adipocyte induction at the tested concentration, while the combination enhanced several differentiation readouts. These are cultured-cell phenotypes.
Literature-Reported Treatment Conditions
- Treatment concentration
- 1, 3 and 10 μM in Figure 2a; 3 μM for Figure 2b–h
- Exposure duration
- 8 days
- Method
- Cell-culture exposure
- Vehicle
- DMSO
- Co-treatment
- RepSox alone and 3 μM RepSox plus 1 μM rosiglitazone were compared with DMSO and 1 μM rosiglitazone alone. Fibroblast growth medium; no MDI induction cocktail.
- Reported limits
- Supplier, lot, analytical purity and explicit salt form were not specified in the read material; n is reported per endpoint without a uniform biological-replicate definition.
Methods and source locations
Compound reported in the paper: RepSox; Compound label in the paper: RepSox (E-616452; SJN 2511)
Published named-compound experiments; salt form and batch identity were not analytically verified for this catalog product.
Figure 2a–h and caption; Results, RepSox induces brown adipocytes from MEFs; Methods, Cell culture and differentiation and endpoint assays
Lipid accumulation and triglyceride content: Oil red O staining and triglyceride assay; mean ± SEM; two-tailed Student’s t-test — Figure 2a–h and caption; Results, RepSox induces brown adipocytes from MEFs; Methods, Cell culture and differentiation and endpoint assays
UCP1 protein and UCP1-positive cells: Western blot with β-tubulin loading control; immunofluorescence; mean ± SEM for quantification — Figure 2a–h and caption; Results, RepSox induces brown adipocytes from MEFs; Methods, Cell culture and differentiation and endpoint assays
Basal, uncoupled and maximal oxygen consumption: Seahorse assay with oligomycin, FCCP and rotenone/antimycin A; mean ± SEM — Figure 2a–h and caption; Results, RepSox induces brown adipocytes from MEFs; Methods, Cell culture and differentiation and endpoint assays
RepSox effects on osteosarcoma cell viability
Cell-based study · Human HOS and 143B osteosarcoma cell lines [1]
RepSox lowered the CCK-8 viability readout with increasing concentration and exposure time. The reported 96-hour IC50 values are cellular assay results, not receptor-binding potency.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0–200 μM
- Exposure duration
- 24, 48 and 96 h
- Method
- Cell-culture exposure
- Vehicle
- DMSO negative control
- Co-treatment
- None
- Reported limits
- DMSO percentage and reagent lot/purity were not specified in the read methods.
Methods and source locations
Compound reported in the paper: RepSox; Compound label in the paper: RepSox (E-616452; SJN 2511)
Published named-compound experiments; salt form and batch identity were not analytically verified for this catalog product.
Figure 1A–B and caption; Methods, Media and reagents, Cell culture, Cell proliferation assays; Results first subsection
HOS CCK-8 viability at 96 h: CCK-8 absorbance at 450 nm; nonlinear concentration-response fitting; Figure 1B and caption — Figure 1A–B and caption; Methods, Media and reagents, Cell culture, Cell proliferation assays; Results first subsection
143B CCK-8 viability at 96 h: CCK-8 absorbance at 450 nm; nonlinear concentration-response fitting; Figure 1B and caption — Figure 1A–B and caption; Methods, Media and reagents, Cell culture, Cell proliferation assays; Results first subsection
JNK and Smad3 phosphorylation in osteosarcoma cells
Cell-based study · Human HOS and 143B osteosarcoma cell lines [1]
RepSox reduced JNK and Smad3 phosphorylation at higher tested concentrations. p38 and ERK phosphorylation were not significantly altered. This supports a cellular signaling association without establishing direct JNK inhibition or pathway causality.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0, 5, 10 and 20 μM (Figure 4B–E labels)
- Method
- Cell-culture exposure
- Vehicle
- Control lane (0 μM); DMSO is the stated solvent
- Co-treatment
- None
- Reported limits
- Exposure duration for Figure 4B–E is not specified in its caption or the shared western-blot method. The 24 h statement for Figure 4A is not transferred to B–E.
Methods and source locations
Compound reported in the paper: RepSox; Compound label in the paper: RepSox (E-616452; SJN 2511)
Published named-compound experiments; salt form and batch identity were not analytically verified for this catalog product.
Figure 4B–E and caption; Methods, Media and reagents and Western blot analysis; Results, RepSox exerts anti-OS effects via the JNK/Smad3 pathway
Phosphorylated/total JNK and Smad3: Western blot densitometry normalized to total JNK or Smad3; GAPDH loading control — Figure 4B–E and caption; Methods, Media and reagents and Western blot analysis; Results, RepSox exerts anti-OS effects via the JNK/Smad3 pathway
Phosphorylated/total p38 and ERK: Western blots and densitometry normalized to corresponding total protein; mean ± SD from three independent experiments — Figure 4B–E and caption; Methods, Media and reagents and Western blot analysis; Results, RepSox exerts anti-OS effects via the JNK/Smad3 pathway
CD34-positive fraction maintained in primary AML/MDS cultures
Cell-based study · Primary human CD34-positive AML/MDS specimens P1–P7; healthy-donor comparator [3]
RepSox reduced the decline in the proportion of CD34-positive cells compared with DMSO. A healthy-donor comparator did not show the same preservation. This phenotype result should not be interpreted as an absolute viable-cell expansion measurement.
Literature-Reported Treatment Conditions
- Treatment concentration
- 16 μM
- Exposure duration
- 6 days
- Method
- Cell-culture exposure
- Vehicle
- DMSO
- Co-treatment
- Bone marrow stromal coculture with 10% FBS and 5% O2; RepSox added once at initiation
- Reported limits
- Reagent lot, purity and salt form were not stated; the aggregate includes AML/MDS specimens rather than a homogeneous AML cohort.
Methods and source locations
Compound reported in the paper: RepSox; Compound label in the paper: RepSox (E-616452; SJN 2511)
Published named-compound experiments; salt form and batch identity were not analytically verified for this catalog product.
Figure 4A–B and caption; Results, RepSox Slows Decay of CD34+ AML/MDS Cells; Methods, Cell Culture, Surface Immunostaining, Flow Cytometric Analysis and Statistical Analysis
Decline in proportion of CD34-positive cells across P1–P7: Flow cytometry with dead-cell exclusion by propidium iodide and matched isotype controls; paired two-tailed Student t-test — Figure 4A–B and caption; Results, RepSox Slows Decay of CD34+ AML/MDS Cells; Methods, Cell Culture, Surface Immunostaining, Flow Cytometric Analysis and Statistical Analysis
CD34-positive fraction in healthy-donor comparator: Flow cytometry; Figure 4A–B and accompanying Results — Figure 4A–B and caption; Results, RepSox Slows Decay of CD34+ AML/MDS Cells; Methods, Cell Culture, Surface Immunostaining, Flow Cytometric Analysis and Statistical Analysis
Concentration-dependent reversal of fibroblast marker responses
Cell-based study · CRL-2097 human dermal fibroblasts [4]
At 100 nM RepSox, basal α-SMA and calponin expression decreased; higher concentrations up to 10 μM instead increased these myofibroblast-associated proteins. The reversal limits a uniform antifibrotic interpretation.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0, 1, 10, 1×10², 1×10³ and 10⁴ nM (Figure 6A labels)
- Exposure duration
- Until day 4
- Method
- Cell-culture exposure
- Co-treatment
- None for Figure 6A
- Reported limits
- Vehicle and replicate count specifically for the Figure 6A blot are not explicit. The 2≤n≤4 statement accompanies the Figure 6 combination quantification and is not assigned to panel A.
Methods and source locations
Compound reported in the paper: RepSox; Compound label in the paper: RepSox (E-616452; SJN 2511)
Published named-compound experiments; salt form and batch identity were not analytically verified for this catalog product.
Figure 6A and caption; Results paragraph beginning Since ERK and JNK inhibition resulted; Methods, Reagents, Cell culture, SDS-PAGE and western blotting
α-SMA and calponin protein expression: Western blot with Histone H3 loading control; representative panel A and Results text — Figure 6A and caption; Results paragraph beginning Since ERK and JNK inhibition resulted; Methods, Reagents, Cell culture, SDS-PAGE and western blotting
