Journal: Anais da Academia Brasileira de Ciencias

Article Title: Huanglian Decoction treats Henoch-Schonlein purpura nephritis by inhibiting NF-κB/NLRP3 signaling pathway and reducing renal IgA deposition.

doi: 10.1590/0001-3765202420220970

Figure Lengend Snippet: Figure 1. Effect of HLD on biochemical indexes of HSPN mice. The levels of total protein (TP), albumin (ALB), blood creatinine (CRE), blood urea (URA), interleukin-1β (IL-1β), interleukin-17 (IL-17), interleukin-18 (IL-18), and tumor necrosis factor-α (TNF-α) in serum of mice. The data are expressed as the mean ± SEM. Compared with the Con group, **P<0.01; compared with the HSPN group, ##P<0.01. Con: control group; HSPN: model group; HSPN+HLD-L: low-dose group of Huanglian Decoction; HSPN+HLD-M: medium-dose group of Huanglian Decoction; HSPN+HLD-H: high-dose group of Huanglian Decoction; HSPN+MMF: mycophenolate mofetil group.

Article Snippet: Then, the Con and HSPN groups were gavaged with distilled water at 0.1 mL each; the HSPN+HLD-L, HSPN+HLD-M and HSPN+HLD-H groups were gavaged with HLD (7.5 g/kg, 15 g/kg and 30 g/kg, respectively) and the HSPN+MMF group was gavaged with MMF (0.3 g/ kg, MedChemExpress, China), the total gavage was 4 weeks.

Techniques: Control

Journal: Anais da Academia Brasileira de Ciencias

Article Title: Huanglian Decoction treats Henoch-Schonlein purpura nephritis by inhibiting NF-κB/NLRP3 signaling pathway and reducing renal IgA deposition.

doi: 10.1590/0001-3765202420220970

Figure Lengend Snippet: Figure 2 Effect of HLD on kidney injury in HSPN mice. (a) Hematoxylin-eosin (HE) staining observation of kidney tissue (400×, Scale bar, 50 μm), black arrow: mesangial cell proliferation, green arrow: vascular congestion. (b) Periodic Acid-Schiff (PAS) staining observation of kidney tissue (400×, Scale bar, 50 μm), red arrow: Positive expression of PAS staining. (c) Immunohistochemical staining was performed to observe renal tissue IgA (20×, scale bar, 50 μm). (d) Immunohistochemical staining was performed to observe renal tissue IgG (20×, scale bar, 50 μm). (e) Immunohistochemical staining was performed to observe renal tissue C3 (20×, scale bar, 50 μm). (f) Percentage of positive expression area. (G) %IgA positive tissue. (h) %IgG positive tissue. (i) %C3 positive tissue. The data are expressed as the mean ± SEM. Compared with the Con group, **P<0.01; compared with the HSPN group, #P<0.05, ##P<0.01; compared with the HSPN+MMF group, ΔP<0.05ΔΔP<0.01.

Article Snippet: Then, the Con and HSPN groups were gavaged with distilled water at 0.1 mL each; the HSPN+HLD-L, HSPN+HLD-M and HSPN+HLD-H groups were gavaged with HLD (7.5 g/kg, 15 g/kg and 30 g/kg, respectively) and the HSPN+MMF group was gavaged with MMF (0.3 g/ kg, MedChemExpress, China), the total gavage was 4 weeks.

Techniques: Staining, Expressing, Immunohistochemical staining

Journal: Anais da Academia Brasileira de Ciencias

Article Title: Huanglian Decoction treats Henoch-Schonlein purpura nephritis by inhibiting NF-κB/NLRP3 signaling pathway and reducing renal IgA deposition.

doi: 10.1590/0001-3765202420220970

Figure Lengend Snippet: Figure 3 Effect of HLD on NF-κB/NLRP3 signaling pathway in the kidney of HSPN mice. (a) Immunofluorescence staining for p65 expression in renal tissues (40×, scale bars, 20 μm). (b) Relative protein expression of IκBα, p-IκBα, NLRP3, caspase-1, and IL-1β, those in western blot assays were expressed after being normalized to β-actin. The data are expressed as the mean ± SEM. Compared with the Con group, **P<0.01; compared with the HSPN group, #P<0.05, ##P<0.01; compared with the HSPN+MMF group, ΔΔP<0.01. DAPI: 4,6-diamidino-2-phenylindole.

Article Snippet: Then, the Con and HSPN groups were gavaged with distilled water at 0.1 mL each; the HSPN+HLD-L, HSPN+HLD-M and HSPN+HLD-H groups were gavaged with HLD (7.5 g/kg, 15 g/kg and 30 g/kg, respectively) and the HSPN+MMF group was gavaged with MMF (0.3 g/ kg, MedChemExpress, China), the total gavage was 4 weeks.

Techniques: Immunofluorescence, Staining, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Early treatment with nootkatone prevents pressure overload-induced ventricular remodeling and heart failure

doi: 10.3389/fphar.2025.1702627

Figure Lengend Snippet: Early NKT administration downregulated the TGF-β1/Smad3 signaling pathway (n = 6–8). (A) mRNA expression of Col Ⅰ and Col Ⅲ in heart tissue. (B) Protein expression of Col Ⅰ, Col Ⅲ, HSP-90, TGF-β1, Smad3, and p-Smad3 in heart tissue. (C) Protein quantitative analysis of Col Ⅰ, Col Ⅲ, HSP-90, TGF-β1, Smad3, and p-Smad3. * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, no significance.

Article Snippet: The membrane was incubated with primary antibodies against collagen type I (Col I, 1:500, ServiceBio, China), collagen type III (Col III, 1:500, ServiceBio, China), HSP-90 (1:2000, OriGene, China), phosphorylated Smad3 (p-Smad3, 1:4000, Wanleibio, China), Smad3 (1:4000, Proteintech, China), transforming growth factor (TGF-β1, 1:5000, Proteintech, China), and β-actin (1:2000, EPITMICS, China), followed by incubation with corresponding HRP-labeled secondary antibodies (ServiceBio, China).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Enhanced cytotoxic effect of radiation and temozolomide in malignant glioma cells: targeting PI3K-AKT-mTOR signaling, HSP90 and histone deacetylases

doi: 10.1186/1471-2407-14-17

Figure Lengend Snippet: Targeting PI3K-Akt-mTOR signaling. (A) U251 and T98G cells were pretreated with rapamycin (RPM) plus TMZ for 24 h and subjected to western blot analysis using the indicated antibodies. Pretreatment with rapamycin (100 nM) plus TMZ (25 μM) did not have a synergistic radiosensitizing effect compared to TMZ alone treatment on U251 and T98G cells. (B) U251 and T98G cells were pretreated with a dual inhibitor of class I PI3K and mTOR signaling, PI103 (0.4 μM), and TMZ (25 μM) for 24 h. PI103 effectively enhanced the radiosensitizing effect of TMZ in both U251 and T98G cells. (C) U251 and T98G cells were pretreated with the HSP90 inhibitor 17-DMAG (25nM) and TMZ (25 μM) for 24 h. 17-DMAG enhanced the radiosensitizing effect of TMZ in U251 and T98G cells. (D) U251 and T98G cells were pretreated with TMZ (25 μM) and LBH589 (20 nM) for 24 h. LBH589 effectively potentiated the radiosensitizing effect of TMZ. Points on survival curves represent mean surviving fractions from minimum three experiments performed in triplicate.

Article Snippet: Membranes were blocked in PBS containing 0.1% Tween 20 and 5% powdered milk and probed with primary antibody directed against p-EGFR (Tyr1068), p-Akt (Ser473), p-ERK (Tyr202/204), p-p70S6K (Thr421/Ser424), HSP70, HSP90, DNA-PKs (Thr2609), Rad51, caspase-3, LC3, MMP-2, E-cadherin, and EphA2 (Cell Signaling Technology, Inc.) at 1:1000 dilutions.

Techniques: Western Blot

Journal: BMC Cancer

Article Title: Enhanced cytotoxic effect of radiation and temozolomide in malignant glioma cells: targeting PI3K-AKT-mTOR signaling, HSP90 and histone deacetylases

doi: 10.1186/1471-2407-14-17

Figure Lengend Snippet: Impairment of DNA damage repair following irradiation. (A) U251 cells were pretreated with the indicated inhibitors plus TMZ before assessment of γH2AX foci formation. Mock-treated control cells were analyzed 6 h after irradiation with 6 Gy. Pretreatment of U251 cells with the dual inhibitor PI103, the HSP90 inhibitor 17-DMAG, or the HDAC inhibitor LBH589 plus TMZ caused marked prolongation of radiation-induced γH2AX foci formation 6 h after 6Gy irradiation. (B) Pretreatment of U251 cells with TMZ combined with PI103, 17-DMAG, or LBH589 attenuated p-DNA-PK expression.

Article Snippet: Membranes were blocked in PBS containing 0.1% Tween 20 and 5% powdered milk and probed with primary antibody directed against p-EGFR (Tyr1068), p-Akt (Ser473), p-ERK (Tyr202/204), p-p70S6K (Thr421/Ser424), HSP70, HSP90, DNA-PKs (Thr2609), Rad51, caspase-3, LC3, MMP-2, E-cadherin, and EphA2 (Cell Signaling Technology, Inc.) at 1:1000 dilutions.

Techniques: Irradiation, Control, Expressing

Journal: Frontiers in Oncology

Article Title: Rationally Repurposing Ruxolitinib ( Jakafi ® ) as a Solid Tumor Therapeutic

doi: 10.3389/fonc.2016.00142

Figure Lengend Snippet: (Ruxolitinib + afatinib) rapidly reduces CDC37 Serine 13 phosphorylation and after 6 h CDC37 total expression . (A,B) SUM149 cells were treated with vehicle control or with (ruxolitinib + afatinib) for 2 h or for 6 h after which cells were fixed in place and permeabilized using 0.5% Triton X100. Immuno-fluorescence was performed: (A) at 10× magnification to detect the total expression and phosphorylation levels of CDC37 and CDC37 Serine 13, respectively; (B) at 60× magnification the total expression of HSP90 and the total expression of CDC37, and the co-localization of the two proteins ( n = 3 ± SEM) * p < 0.05 greater decline than CDC37 protein expression. (C) SUM149 cells were transfected with empty vector control or with plasmids to express: HSP90, GRP78, HSP70, or HSP27, alone or in the indicated combinations. Twenty-four hours after transfection, cells were treated with vehicle control or with [ruxolitinib (1.0 μM) + afatinib (1.0 μM)] for 6 h after which cells were fixed in place and permeabilized using 0.5% Triton X100. Immuno-fluorescence was performed at 10× magnification to detect the total expression of MCL-1 and BCL-XL ( n = 3 ± SEM) * p < 0.05 lower decline than vehicle control. (D) SUM149 cells were transfected with empty vector control or with plasmids to express: HSP90, GRP78, HSP70, or HSP27, alone or in the indicated combinations. Twenty-four hours after transfection, cells were treated with vehicle control or with [ruxolitinib (1.0 μM) + afatinib (1.0 μM)] for 24 h. Twenty-four hours later, cell viability was assessed using a live/dead assay in a Hermes WiScan microscope at 10× magnification ( n = 3 ± SEM) * p < 0.05 greater survival than individual expression of chaperones.

Article Snippet: Antibodies used include: HSP90 (E289) (Cell Signaling); HSP90 (#2928) (Abcam); HSP90 (ab195575) Abcam; HSP90 3G3 (13495) (Abcam); GRP78 (50b12) (31772) (Cell Signaling); GRP78 (ab191023) Abcam; GRP78 (ab103336) Abcam; GRP78 (N-20) (sc-1050) Santa Cruz; HSP27 (G31) (2402P) Cell Signaling); HSP27 (EP1724Y) (ab62339) Abcam; HSP27 (H-77) (sc-9012) Santa Cruz; HSP27 (LS-C31836) Lifespan science Corp. Other antibodies were as used in prior studies by the laboratory.

Techniques: Expressing, Fluorescence, Transfection, Plasmid Preparation, Live Dead Assay, Microscopy

Journal: Molecular Cancer Therapeutics

Article Title: Cell Intrinsic Role of COX-2 in Pancreatic Cancer Development

doi: 10.1158/1535-7163.mct-12-0342

Figure Lengend Snippet: Figure 7. Decreased FKBP5 negative feedback leads to enhanced AKT activation in Pdx1-Creþ;K-rasG12D/þ;Ptenlox/þ; Cox-2lox/lox mice. A, FKBP5 mRNA expression in PDACs of Pdx1-Creþ; K-rasG12D/þ;Ptenlox/þ;Cox-2lox/lox

Article Snippet: The following primary antibodies were used: phospho-AKT(Ser473) (Cell Signaling; 1:50), Cytokeratin 19 (ab15463, Abcam; 1:100), COX-2 (SP21; Thermo Scientific, ready-to-use),GRP78(11587-1AP,ProteinTechGroup; 1:50), and FKBP5 (14155-1-AP, ProteinTech Group, 1:50).

Techniques: Activation Assay, Expressing

Journal: Scientific Reports

Article Title: Therapeutic effect of histone deacetylase 6 inhibitor for a mouse model of phenylketonuria

doi: 10.1038/s41598-025-29143-7

Figure Lengend Snippet: Western blot analysis of acetyl-HSP90, HSP90, and PAH protein expression in liver tissues from sham, treated, and wild-type groups following 12 weeks of TSA treatment. ( A ) Expression of acetyl-HSP90, HSP90, PAH, and GAPDH (used as a loading control). ( B ) Quantitative analysis of acetyl-HSP90 relative to HSP90 protein levels. ( C ) Quantitative analysis of PAH relative to GAPDH protein levels. The expression of PAE was quantified using the Image J software (Version 1.49) and expressed as arbitrary units (a.u.). Results are presented as mean ± SEM, with statistical significance indicated. ****P<0.0001.

Article Snippet: The antibodies used anti-acetylated HSP90 (600-401-981, Rockland Immunochemical, 1:500), anti-HSP90 (#4877, Cell Signaling, 1:1,000), and PAH (sc-271258, Santa Cruz, 1:2,000).

Techniques: Western Blot, Expressing, Control, Software

Journal: Scientific Reports

Article Title: Therapeutic effect of histone deacetylase 6 inhibitor for a mouse model of phenylketonuria

doi: 10.1038/s41598-025-29143-7

Figure Lengend Snippet: Mechanism of TSA in enhancing HSP90 acetylation and stabilizing misfolded PAH protein to improve phenylalanine metabolism.

Article Snippet: The antibodies used anti-acetylated HSP90 (600-401-981, Rockland Immunochemical, 1:500), anti-HSP90 (#4877, Cell Signaling, 1:1,000), and PAH (sc-271258, Santa Cruz, 1:2,000).

Techniques:

Journal: FEBS Open Bio

Article Title: In silico identification and biochemical characterization of the human dicarboxylate clamp TPR protein interaction network

doi: 10.1002/2211-5463.12521

Figure Lengend Snippet: Graphical representation of human proteins with Hsp70/Hsp90‐like C termini identified by global search of the NCBI human protein database. Schematic diagram of interaction network between proteins with Hsp70/Hsp90‐like C termini (in squares) and dc TPR (in circles) proteins. The connections between various proteins are shown by lines. Grey lines show confirmed interactions of proteins other than Hsp70/Hsp90 via dicarboxylate clamp mechanism. Red lines represent connections between proteins via dicarboxylate clamp mechanism established in this study. Blue lines represent protein interactions within network identified by Taipale et al . . Yellow lines show interactions extracted from protein interaction database ( https://www.ncbi.nlm.nih.gov/gene ). For simplicity of presentation, we did not show interactions between dc TPR proteins and members of Hsp70 and Hsp90 family.

Article Snippet: The following antibodies were used: AIP (SAB2105326) and Nicotinamide adenine dinucleotide synthase 1 (NADSYN1) (WH0055191M1) were from Sigma‐Aldrich, FKBP51 (PA1‐020), mouse monoclonal anti‐AIP (MA3‐16515) and p23 (MA3‐414) were from Thermo Fisher Scientific (Uppsala, Sweden), Usp19 (ab68527), dynein axonemal assembly factor 2 (DNAAF2; ab99056) and Hsp105 (ab74174) were from Abcam (Cambridge, MA, USA), Tom 20 (sc‐11415), Tom70 (sc‐390545), Bcl‐2‐like protein 14 (Bcl‐G) (sc‐398223) and mouse monoclonal anti‐FKBP51 (sc‐271547) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA), and Hsp90 (MAB3286) was from R&D Systems (Minneapolis, MN, USA).

Techniques:

Journal: FEBS Open Bio

Article Title: In silico identification and biochemical characterization of the human dicarboxylate clamp TPR protein interaction network

doi: 10.1002/2211-5463.12521

Figure Lengend Snippet: Analysis of interaction of FKBP 51 and AIP with 10‐amino‐acid peptides corresponding to C termini of Hsp90, Hsp70, Hsp105, Tom20, and p23. (A) Dot‐blot competition assay of interactions between FKBP 51 and Hsp90 peptide conjugated to HRP incubated with increasing amounts of unconjugated Hsp90 peptide. (B) Inhibition curves of FKBP 51 interactions with HRP ‐conjugated 10‐amino‐acid peptides corresponding to C termini of Hsp90, Hsp70, Hsp105, Tom20, and p23 probed with increasing concentrations of label‐free peptides. (C) As in (B) but AIP was used instead of FKBP 51. n = 3. Graphs were created and SD values calculated using prism , version 7.02.

Article Snippet: The following antibodies were used: AIP (SAB2105326) and Nicotinamide adenine dinucleotide synthase 1 (NADSYN1) (WH0055191M1) were from Sigma‐Aldrich, FKBP51 (PA1‐020), mouse monoclonal anti‐AIP (MA3‐16515) and p23 (MA3‐414) were from Thermo Fisher Scientific (Uppsala, Sweden), Usp19 (ab68527), dynein axonemal assembly factor 2 (DNAAF2; ab99056) and Hsp105 (ab74174) were from Abcam (Cambridge, MA, USA), Tom 20 (sc‐11415), Tom70 (sc‐390545), Bcl‐2‐like protein 14 (Bcl‐G) (sc‐398223) and mouse monoclonal anti‐FKBP51 (sc‐271547) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA), and Hsp90 (MAB3286) was from R&D Systems (Minneapolis, MN, USA).

Techniques: Dot Blot, Competitive Binding Assay, Incubation, Inhibition

Journal: FEBS Open Bio

Article Title: In silico identification and biochemical characterization of the human dicarboxylate clamp TPR protein interaction network

doi: 10.1002/2211-5463.12521

Figure Lengend Snippet: In vitro and in situ analysis of interactions of 10‐amino‐acid peptides corresponding to C termini of Hsp90 and Tom20 with Tom70. (A) Dot‐blot assay of interactions between Tom70 and HRP ‐conjugated 10‐amino‐acid peptides corresponding to C termini of Hsp90 and Tom20 in the presence of 20 μ m of unconjugated peptides. (B) Fluorescence imaging of HEK 293 cells in the presence of 20 μ m of fluorescein‐labeled Hsp90, R9–Hsp90, Tom20 and R9–Tom20 peptides. (C) Inhibition of Tom20–Tom70 and Hsp90–Tom70 interactions with R9–Hsp90 and R9–Tom20 in HEK 293 cells measured by PLA. n = 3. Graphs were created and SD values were calculated using prism , version 7.02. * P ‐value < 0.05 compared to control; comparisons are based on one‐way ANOVA . (D) Inhibition of Tom20–Tom70 and Hsp90–Tom70 interactions with R9–Hsp90 and R9–Tom20 in HEK 293 cells measured by mammalian two‐hybrid assay. n = 3. Graphs were created and SD values were calculated using prism , version 7.02. * P ‐value < 0.05 compared to control; comparisons are based on one‐way ANOVA .

Article Snippet: The following antibodies were used: AIP (SAB2105326) and Nicotinamide adenine dinucleotide synthase 1 (NADSYN1) (WH0055191M1) were from Sigma‐Aldrich, FKBP51 (PA1‐020), mouse monoclonal anti‐AIP (MA3‐16515) and p23 (MA3‐414) were from Thermo Fisher Scientific (Uppsala, Sweden), Usp19 (ab68527), dynein axonemal assembly factor 2 (DNAAF2; ab99056) and Hsp105 (ab74174) were from Abcam (Cambridge, MA, USA), Tom 20 (sc‐11415), Tom70 (sc‐390545), Bcl‐2‐like protein 14 (Bcl‐G) (sc‐398223) and mouse monoclonal anti‐FKBP51 (sc‐271547) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA), and Hsp90 (MAB3286) was from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vitro, In Situ, Dot Blot, Fluorescence, Imaging, Labeling, Inhibition, Control, Two Hybrid Assay