Journal: Phytotherapy research : PTR

Article Title: Oleanolic Acid Up-Regulated UGT1A1 and Antagonized Inflammation by Affecting the Binding of PXR and PKCα to HSP90α and SRC1.

doi: 10.1002/ptr.8515

Figure Lengend Snippet: FIGURE 1 | Effects of OA (20 μM) on the expression of UGT1A1 (A, B), HSP90α (B), PKCα (C), PXR (D) and SRC1 (E) in HepG2 cells. The data are presented as the mean ± SD of triplicate independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 compare to control group; #p < 0.05 and

Article Snippet: Primary antibodies against human UGT1A1 (rabbit- monoclonal, ab170858, Lot: GR299245- 5) and rat UGT1A1 (rabbit- polyclonal, ab194697, Lot: GR3209781- 1) were provided by Abcam (Cambridge, MA, USA), PXR (rabbit- polyclonal, A1583, Lot: 3515316101) were obtained from ABclonal (Wuhan, China), and PKCα (rabbit- polyclonal, bs- 3531R, Lot: BB06154399), HSP90α (rabbit- polyclonal, bs10100R, Lot: AH06203629) were bought from Bioss (Beijing, China), while SRC1 (rabbit- polyclonal, 21952- 1- AP, Lot: 00121428), Lamin B1 (rabbit- polyclonal, 66095- 1- Ig, Lot: 10026670), and GAPDH (mouse- monoclonal, Lot: 60004- 1- Ig) were purchased from Proteintech Group Inc. (Wuhan, China).

Techniques: Expressing, Control

Journal: Phytotherapy research : PTR

Article Title: Oleanolic Acid Up-Regulated UGT1A1 and Antagonized Inflammation by Affecting the Binding of PXR and PKCα to HSP90α and SRC1.

doi: 10.1002/ptr.8515

Figure Lengend Snippet: FIGURE 2 | Effects of OA (20 μM) on the PKCα pull-down protein expression of HSP90α (A, B), PXR pull-down protein expression of SRC1 (C, D) and HSP90α (C, E) in HepG2 cells. The data are presented as the mean ± SD of triplicate independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 compare to control group; #p < 0.05 and ##p < 0.01 compare to PMA treated group, respectively.

Article Snippet: Primary antibodies against human UGT1A1 (rabbit- monoclonal, ab170858, Lot: GR299245- 5) and rat UGT1A1 (rabbit- polyclonal, ab194697, Lot: GR3209781- 1) were provided by Abcam (Cambridge, MA, USA), PXR (rabbit- polyclonal, A1583, Lot: 3515316101) were obtained from ABclonal (Wuhan, China), and PKCα (rabbit- polyclonal, bs- 3531R, Lot: BB06154399), HSP90α (rabbit- polyclonal, bs10100R, Lot: AH06203629) were bought from Bioss (Beijing, China), while SRC1 (rabbit- polyclonal, 21952- 1- AP, Lot: 00121428), Lamin B1 (rabbit- polyclonal, 66095- 1- Ig, Lot: 10026670), and GAPDH (mouse- monoclonal, Lot: 60004- 1- Ig) were purchased from Proteintech Group Inc. (Wuhan, China).

Techniques: Expressing, Control

Journal: Phytotherapy research : PTR

Article Title: Oleanolic Acid Up-Regulated UGT1A1 and Antagonized Inflammation by Affecting the Binding of PXR and PKCα to HSP90α and SRC1.

doi: 10.1002/ptr.8515

Figure Lengend Snippet: FIGURE 3 | Effects of OA (20 μM) on the localization of PKCα and HSP90α (A), PXR and HSP90α (B), PXR and SRC1 (C) in HepG2 cells and the structure of compound OA (D).

Article Snippet: Primary antibodies against human UGT1A1 (rabbit- monoclonal, ab170858, Lot: GR299245- 5) and rat UGT1A1 (rabbit- polyclonal, ab194697, Lot: GR3209781- 1) were provided by Abcam (Cambridge, MA, USA), PXR (rabbit- polyclonal, A1583, Lot: 3515316101) were obtained from ABclonal (Wuhan, China), and PKCα (rabbit- polyclonal, bs- 3531R, Lot: BB06154399), HSP90α (rabbit- polyclonal, bs10100R, Lot: AH06203629) were bought from Bioss (Beijing, China), while SRC1 (rabbit- polyclonal, 21952- 1- AP, Lot: 00121428), Lamin B1 (rabbit- polyclonal, 66095- 1- Ig, Lot: 10026670), and GAPDH (mouse- monoclonal, Lot: 60004- 1- Ig) were purchased from Proteintech Group Inc. (Wuhan, China).

Techniques:

Journal: Phytotherapy research : PTR

Article Title: Oleanolic Acid Up-Regulated UGT1A1 and Antagonized Inflammation by Affecting the Binding of PXR and PKCα to HSP90α and SRC1.

doi: 10.1002/ptr.8515

Figure Lengend Snippet: FIGURE 6 | Effects of OA (20 μM) on the PKCα pull-down protein expression of HSP90α (A, B), PXR pull-down protein expression of SRC1 (C, D) and HSP90α (C, E) in HepG2 cells overexpressing hPKCα. The data are presented as the mean ± SD of triplicate independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 compare to control group; #p < 0.05, ##p < 0.01 and ###p < 0.001 compare to hPKCα OE group, respectively.

Article Snippet: Primary antibodies against human UGT1A1 (rabbit- monoclonal, ab170858, Lot: GR299245- 5) and rat UGT1A1 (rabbit- polyclonal, ab194697, Lot: GR3209781- 1) were provided by Abcam (Cambridge, MA, USA), PXR (rabbit- polyclonal, A1583, Lot: 3515316101) were obtained from ABclonal (Wuhan, China), and PKCα (rabbit- polyclonal, bs- 3531R, Lot: BB06154399), HSP90α (rabbit- polyclonal, bs10100R, Lot: AH06203629) were bought from Bioss (Beijing, China), while SRC1 (rabbit- polyclonal, 21952- 1- AP, Lot: 00121428), Lamin B1 (rabbit- polyclonal, 66095- 1- Ig, Lot: 10026670), and GAPDH (mouse- monoclonal, Lot: 60004- 1- Ig) were purchased from Proteintech Group Inc. (Wuhan, China).

Techniques: Expressing, Control

Journal: Frontiers in Endocrinology

Article Title: Cadmium activation of wild-type and constitutively active estrogen receptor alpha

doi: 10.3389/fendo.2024.1380047

Figure Lengend Snippet: Effects of cadmium on ERα wild-type and calcium/metal interaction site mutants in HEK293T cells. HEK293T cells were transfected with wild-type ERα and mutants E542A/D545A, N532A, H516A/N519A/E523A, and C381A. Following transfection, the cells were treated with estradiol (100 nM) or cadmium (2 µM) for 60 minutes, crosslinked, lysed, and the occupancy of ERα on the enhancers of target genes was examined by ChIP assay. A Re-ChIP assay was performed, and the chromatin was first immunoprecipitated with an antibody to ERα, followed by immunoprecipitation with an antibody to SRC1, PolII, or IgG. DNA was quantified by real time qPCR and the recruitment of ERα, SRC1, and PolII is presented as percent input. The data are the mean of two independent experiments done in duplicate (mean ± SEM). (A, B) ChIP wtERα. (C, D) ChIP E542A/D545A. (E, F) ChIP N532A. (G, H) ChIP H516A/N519A/E523A. (I, J) ChIP C381A. (K, L) Re-ChIP wtERα. (M, N) Re-ChIP E542A/D545A. (O, P) Re-ChIP N532A. (Q, R) Re-ChIP H516A/N519A/E523A. (S, T) Re-ChIP C381A.

Article Snippet: For re-ChIP, the second immunoprecipitation was performed with antibodies to SRC1 (Abcam Cat#ab2859, RRID: AB_303360) or PolII (Thermo Fisher Scientific Cat#MA1-26249, RRID: AB_795353).

Techniques: Transfection, Immunoprecipitation

Journal: Frontiers in Endocrinology

Article Title: Cadmium activation of wild-type and constitutively active estrogen receptor alpha

doi: 10.3389/fendo.2024.1380047

Figure Lengend Snippet: Effects of cadmium on the recruitment of Y537S and Y537S calcium/metal interaction site mutants to the enhancers of estrogen responsive genes. HEK293T cells were transfected with Y537S ERα and mutants Y537S/E542A/D545A, Y537S/N532A, Y537S/H516A/N519A/E523A, and Y537S/C381A. Following transfection, the cells were treated with estradiol (100 nM) or cadmium (2 µM) for 60 minutes, crosslinked, lysed, and the occupancy of ERα on the enhancers of target genes was examined by ChIP assay. A Re-ChIP assay was performed, and the chromatin was first immunoprecipitated with an antibody to ERα, followed by immunoprecipitation with an antibody to SRC1, PolII, or IgG. DNA was quantified by real time qPCR and the recruitment of ERα, SRC1, and PolII is presented as percent input. The data are the mean of two independent experiments done in duplicate (mean ± SEM). (A, B) ChIP Y537S. (C, D) ChIP Y537S/E542A/D545A. (E, F) ChIP Y537S/N532A. (G, H) ChIP Y537S/H516A/N519A/E523A. (I, J) ChIP Y537S/C381A. (K, L) Re-ChIP Y537S. (M, N) Re-ChIP Y537S/E542A/D545A. (O, P) Re-ChIP Y537S/N532A. (Q, R) Re-ChIP Y537S/H516A/N519A/E523A. (S, T) Re-ChIP Y537S/C381A.

Article Snippet: For re-ChIP, the second immunoprecipitation was performed with antibodies to SRC1 (Abcam Cat#ab2859, RRID: AB_303360) or PolII (Thermo Fisher Scientific Cat#MA1-26249, RRID: AB_795353).

Techniques: Transfection, Immunoprecipitation

Journal: Frontiers in Endocrinology

Article Title: Cadmium activation of wild-type and constitutively active estrogen receptor alpha

doi: 10.3389/fendo.2024.1380047

Figure Lengend Snippet: Effects of cadmium on the recruitment of D538G and D538G calcium/metal interaction site mutants to the enhancers of estrogen responsive genes. HEK293T cells were transfected with D538G ERα and mutants D538G/E542A/D545A, D538G/N532A, D538G/H516A/N519A/E523A, and D538G/C381A. Following transfection, the cells were treated with estradiol (100 nM) or cadmium (2 µM) for 60 minutes, crosslinked, lysed, and the occupancy of ERα on the enhancers of target genes was examined by ChIP assay. A Re-ChIP assay was performed, and the chromatin was first immunoprecipitated with an antibody to ERα, followed by immunoprecipitation with an antibody to SRC1, PolII, or IgG. DNA was quantified by real time qPCR and the recruitment of ERα, SRC1, and PolII is presented as percent input. The data are the mean of two independent experiments done in duplicate (mean ± SEM). (A, B) ChIP D538G. (C, D) ChIP D538G/E542A/D545A. (E, F) ChIP D538G/N532A. (G, H) ChIP D538G/H516A/N519A/E523A. (I, J) ChIP D538G/C381A. (K, L) Re-ChIP D538G. (M, N) Re-ChIP D538G/E542A/D545A. (O, P) Re-ChIP D538G/N532A. (Q, R) Re-ChIP D538G/H516A/N519A/E523A. (S, T) Re-ChIP D538G/C381A.

Article Snippet: For re-ChIP, the second immunoprecipitation was performed with antibodies to SRC1 (Abcam Cat#ab2859, RRID: AB_303360) or PolII (Thermo Fisher Scientific Cat#MA1-26249, RRID: AB_795353).

Techniques: Transfection, Immunoprecipitation

Journal: Molecular cell

Article Title: UBR5 forms ligand-dependent complexes on chromatin to regulate nuclear hormone receptor stability

doi: 10.1016/j.molcel.2023.06.028

Figure Lengend Snippet:

Article Snippet: Rabbit anti-SRC1 , CST , Clone: 128E7; RRID:AB_2196189.

Techniques: Virus, Recombinant, Protease Inhibitor, Ubiquitin Proteomics, Staining, Software, Expressing