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: 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: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: USP50 stimulates HMGB1 release in DGR-induced gastric inflammation. (A, B) mRNA levels of HMGB1, HSP70, HSP90, S100A8, and S100A9 were detected via qRT-PCR in U937 cells (A) or RAW264.7 cells (B) treated with DMSO or TDCA (200μM, 24 h). (C-H) ELISA was used to measure the concentrations of HMGB1 (C) , HSP70 (D) , HSP90 (E) , S100A8 (F) , and S100A9 (G) in the supernatant of U937 cells and RAW264.7 cells, while ATP concentration (H) was surveyed by ATP assay kit. (I) The protein level of HMGB1 was detected in gastric tissues of Sham and GJ mice by western blotting. Image J was used for quantitative analysis, with GAPDH as loading control. (J) The level of HMGB1 between normal gastric tissues and gastric cancer tissues was analyzed via GEPIA database (gepia.cancer-pku.cn/). (K) The IHC staining of HMGB1 was performed with sections from BRG patients and GC patients. (L) IRS was calculated for (K) . (M, N) HMGB1 was assayed by ELISA in the supernatant of U937 cells with USP50 overexpression (M) or knock-down (N) . * P < 0.05, ** P < 0.01, and *** P <0.001. "ns" means "no significance" ( P >0.05).

Article Snippet: Human S100A8 (E-EL-H1289c), Human S100A9 (E-EL-H1290c), HSP90 ELISA Kit (E-EL-H1864c), Human HMGB-1 ELISA Kit (E-EL-H1554c), Mouse HMGB-1 ELISA Kit (E-EL-M0676c) were obtained from Elabscience Biotechnology Co., Ltd, China.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Concentration Assay, ATP Assay, Western Blot, Control, Immunohistochemistry, Over Expression, Knockdown

Journal: Frontiers in Physiology

Article Title: Does Predation Exacerbate the Risk of Endosymbiont Loss in Heat Stressed Hermatypic Corals? Molecular Cues Provide Insights Into Species-Specific Health Outcomes in a Multi-Stressor Ocean

doi: 10.3389/fphys.2022.801672

Figure Lengend Snippet: Factorial MANOVAs and ANOVAs testing the main and interactive effects of (i) species, (ii) predation, and (iii) temperature in biomarkers Hsp70, Ub, and TAC concentrations: (A) for coral species that survived up to 30°C (all seven species); (B) for coral species that also survived at 32°C (thermotolerant Galaxea fascicularis , Psammocora contigua, and Turbinaria reniformis ).

Article Snippet: Hsp70 and Ub concentrations were then calculated using calibration curves constructed from serial dilutions of purified Hsp70 active protein (OriGene, Germany) and of purified Ub (UbpBIO, United States) standards, respectively, to give a range of 0.0 to 2.0 mg ml −1 .

Techniques:

Journal: Cell death and differentiation

Article Title: Combined absence of TRP53 target genes ZMAT3, PUMA and p21 cause a high incidence of cancer in mice.

doi: 10.1038/s41418-023-01250-w

Figure Lengend Snippet: Fig. 3 The combined absence of ZMAT3 and p21 does not accelerate γ-radiation induced thymic lymphoma development. A Kaplan- Meier curves showing percentages of tumour-free mice of the indicated genotypes after exposure to four weekly doses of γ-radiation (1.5 Gy for each dose). Differences in thymic lymphoma incidence between wt and p21−/−Zmat3−/−were not statistically significant. P value determined by log-rank (Mantel-Cox) test p = 0.5. B Immunophenotyping of γ-radiation-induced thymic lymphomas arising in mice of the indicated genotypes, as assessed by cell surface marker staining and flow cytometric analysis of tumour cells from the thymus. Data are presented as the frequency of the indicated phenotypes for each genotype. Double negative CD4-CD8- (DN) or double positive CD4+CD8+ (DP) lymphoma cells found in the thymus that show a trend towards the CD8 T cell lineage (DP/CD8+ or DN CD8+, respectively). N number of mice. C Thymus weights from sick mice of the indicated genotypes. Significant differences were observed in spleen weights between the sick p21−/−Zmat3−/−and sick wt γ-irradiated mice. p21−/−Zmat3−/−(N = 12), p21−/−(N = 7), Zmat3−/−(N = 7) and wt (N = 5). Mean ± SEM, Unpaired Students t-test **p = 0.0065. D Western blot analysis of p19ARF, TRP53 and HSP70 (loading control) in thymic lymphomas of the indicated genotypes. Red asterisk indicates samples with dysregulated TRP53 pathway. The Trp53 mutant Eμ-Myc lymphoma cell line EMRK1172 [51] was used as a control for p19ARF and mutant TRP53 protein overexpression. Protein size standards in kilodaltons (kDa) are indicated. The numbers represent the identification of individual mice in the colony. E Trp53 exons 4-11 were sequenced from tumours (N = 26, Table S4). Table summary of mutations found, listed by amino acid change, Trp53 knock out (KO) or Trp53 wild-type (WT) alleles. F Summary of the TRP53 status in γ-radiation-induced thymic lymphomas arising in mice of the indicated genotypes as assessed by Western blot analysis and/or Trp53 exon sequencing (see also Table S4). Inconclusive refers to lymphomas where TRP53 pathway dysregulation was seen by only Western blot analysis or exon sequencing but not in both assays.

Article Snippet: The following antibodies were used for probing; mouse TRP53 (clone CM5, Novocastra), p19/ARF (clone 5.C3.1, Rockland) and HSP70 (clone N6, gift from Dr. R Anderson, Olivia Newton-John Cancer Research Institute, Melbourne, VIC, Australia), the latter used as a control for protein loading.

Techniques: Marker, Staining, Irradiation, Western Blot, Control, Mutagenesis, Over Expression, Knock-Out, Sequencing