In HepG2 cells, 800 nM U0126 increased CYP7A1 mRNA after 0.5 h pretreatment and an additional 10 h treatment. In a separate EA.hy926 experiment using A10957, a condition labeled MEK1/2 inhibitors prevented TNFα-induced ERK1/2 phosphorylation while CO2-induced activation persisted. Its caption specifies U0126 (2.5 µM) and PD98059 (25 µM), without separate single-agent responses, so the latter effects cannot be assigned to U0126 alone. [1,2]
U0126 alters ERK phosphorylation and CYP7A1 expression in HepG2 cells
Cell-based study · Human HepG2 hepatoma cells [1]
At 800 nM, U0126 reduced phospho-ERK1/2 signal and increased CYP7A1 mRNA relative to the U0126-negative condition. This is one cell-treatment design with two related endpoints, not direct inhibition of isolated ERK enzyme.
Literature-Reported Treatment Conditions
- Treatment concentration
- 800 nM
- Exposure duration
- 0.5 h pretreatment followed by an additional 10 h treatment (Fig. 6d,e caption)
- Method
- HepG2 cell treatment; cells serum-starved for 10 h before subsequent treatment in shared methods.
- Co-treatment
- No co-treatment is indicated for the U0126 +/- comparison in Fig. 6d,e.
- Reported limits
- U0126 vehicle and exact biological n for Fig. 6d,e are not specified in the acquired text/caption. Global statement: experiments repeated at least twice; qPCR triplicate analyses are technical replicates, not a biological n.
Methods and source locations
Compound reported in the paper: U0126; Compound label in the paper: Adooq Bioscience A10957
Evidence for the paper-reported A10957 reagent; no current lot, purity, stereochemical or solvate validation and no salt/form conversion.
Fig. 6d,e (image and caption); Materials and methods: Reagents, Cell culture and treatment, Quantitative real-time PCR, Western blotting, Statistical analysis; Results: FGF21-induced ERK activity mediated the Cyp7a1 induction by BCAAs
ERK1/2 phosphorylation: Western blot for phospho-ERK1/2, total ERK1/2 and β-actin — Fig. 6d; Western blotting
CYP7A1 mRNA expression: qPCR normalized to 18S RNA; mean ± SEM; two-sided Student t-test for two-group comparisons — Fig. 6e; Quantitative real-time PCR; Statistical analysis
Stimulus-dependent ERK phosphorylation under the reported MEK-inhibitor condition
Cell-based study · Human EA.hy926 endothelial cell line [2]
The condition labeled MEK1/2 inhibitors prevented TNFα-induced ERK1/2 phosphorylation, while a CO2-induced increase persisted. The caption lists U0126 and PD98059, without separate single-agent responses; the effect cannot be assigned to U0126 alone.
Literature-Reported Treatment Conditions
- Treatment concentration
- U0126 2.5 µM; PD98059 25 µM
- Exposure duration
- 0 and 5 min stimulus time points in Fig. 1e; inhibitor preincubation time is not specified for this panel.
- Method
- EA.hy926 cells exposed to 10% CO2 or 10 ng/mL TNFα under the MEK1/2-inhibitor condition.
- Vehicle
- PBS and/or DMSO controls specified in shared cell-culture methods; exact panel-specific fraction not stated.
- Co-treatment
- Fig. 1e shows one condition labeled MEK1/2 inhibitors and the caption specifies U0126 2.5 µM and PD98059 25 µM. No separate single-agent lanes are identifiable; simultaneous versus otherwise grouped use is not explicitly distinguished in the selected text.
- Reported limits
- Cannot isolate the contribution of U0126 from PD98059 or establish simultaneous versus grouped use from the panel. Inhibitor preincubation time and panel-specific vehicle fraction are not specified in the acquired selected text.
Methods and source locations
Compound reported in the paper: U0126; Compound label in the paper: Adooq Bioscience A10957
Evidence for the paper-reported A10957 reagent; no current lot, purity, stereochemical or solvate validation and no salt/form conversion.
Fig. 1e (image, condition labels, full caption); Materials and methods: Cell culture, Immunoblotting, Statistical analysis; Results: ERK1/2 are activated by CO2 in vitro and in human cells
Phospho-ERK1/2 response to TNFα versus CO2: Phospho-ERK1/2 immunoblotting and densitometry; total ERK1/2, GAPDH and Ponceau S loading controls; mean ± SD of three independent experiments — Fig. 1e; full Fig. 1 caption; Statistical analysis
