Hydrocortisone concentration response of GR phosphorylation and localization
Cell-based study · HEK 293T cells transfected with FLAG-GR and GFP-FKBP51 (1:1 plasmid ratio) [1]
After 24 h, hydrocortisone increased GR Ser211 phosphorylation and shifted GR distribution toward the nucleus. The nuclear response was weaker than the cytoplasmic response. These are cellular signaling endpoints, without a receptor-binding affinity or an EC50 determination. Figure caption reports n = 3 and mean ± SEM; the experimental unit underlying n is not specified.
| Endpoint | Reported result | Reference |
|---|---|---|
| GR Ser211 phosphorylation relative to total GR | One-way ANOVA; caption: mean ± SEM, n = 3 Concentration-dependent increase; F(5,12) = 60.66, p < 0.0001, η² = 0.96 | [1] |
| Cytoplasmic GR relative to α-tubulin | One-way ANOVA; caption: mean ± SEM, n = 3 Decrease with increasing concentration; F(5,12) = 21.93, p < 0.0001, η² = 0.90 | [1] |
| Nuclear GR relative to histone H3 | One-way ANOVA; caption: mean ± SEM, n = 3 Increase across concentration groups; F(5,12) = 3.20, p = 0.046, η² = 0.57 | [1] |
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.1, 1, 10, 100 and 1000 nM
- Exposure duration
- 24 h
- Method
- Added to transfected cell cultures; Western blot of whole-cell and fractionated proteins
- Vehicle
- DMSO; equal-volume vehicle control
- Reported limits
- Final DMSO percentage and biological versus technical meaning of n = 3 are not specified in the relevant methods/caption. No Adooq lot was tested.
Methods and source locations
Compound reported in the paper: Hydrocortisone (cortisol); Compound label in the paper: Tocris catalog 4093
Evidence concerns hydrocortisone from another supplier in a transfected cell system; it does not validate Adooq lot performance or animal efficacy.
Methods 2.4–2.7; Results 3.3; Figure 5A,C,D and caption (both Figure 5 images viewed)
GR Ser211 phosphorylation relative to total GR: Whole-cell Western blot; pGR Ser211/GR ratio normalized to DMSO vehicle — Figure 5A; Results 3.3
Cytoplasmic GR relative to α-tubulin: Fractionated-cell Western blot; GR/α-tubulin normalized to DMSO vehicle — Figure 5C; Results 3.3
Nuclear GR relative to histone H3: Fractionated-cell Western blot; GR/histone H3 normalized to DMSO vehicle — Figure 5D; Results 3.3