Journal: PLOS One

Article Title: Mitochondria-targeted ROS scavenger JP4-039 improves cardiac function in a post-myocardial infarction animal model and induces angiogenesis in vitro

doi: 10.1371/journal.pone.0320703

Figure Lengend Snippet: In acute MI, the exacerbation of Mito-ROS leads to mitochondrial dysfunction and a decrease in ATP production, leading to an increase in AMP/ATP ratio, which results in phosphorylation of AMPKα at Threonine 172. Over time, the unmet need for ATP results in impaired angiogenesis and loss of capillaries in the heart, affecting blood supply and myocardial healing. This leads to cardiac maladaptation, resulting in impaired cardiac function associated with larger infarcted tissue area after acute MI. When JP4-039 is introduced after acute MI, its mitochondria-targeted ROS and electron scavenging activities result in a reduction of Mito-ROS and increased expression of mitochondrial complexes I and V. Thus, treatment with JP4-039 results in increased ATP supply and reduced phosphorylation of AMPKα at Threonine 172 in comparison to untreated mice after MI, suggesting an improvement of the AMP/ATP balance. The increase in ATP supply promoted by JP4-039 may contribute to angiogenesis associated with an improved cardiac function and reduction of infarct size after an acute MI. Created with BioRender.com.

Article Snippet: Human coronary artery endothelial cells (HCAEC), or MHEC with prolonged overexpression of NOX2 were plated into a 15-well µ-slide Angiogenesis (ibidi GmbH, Gräfelfing, Germany) precoated with Cultrex® basement membrane extract (BME, R&D Systems, Minneapolis, MN) at a density of 10 4 cells/well (passage 4), following manufacturer protocol [ ].

Techniques: Expressing, Comparison

Journal: Molecular Therapy

Article Title: The role of the mitochondrial protein VDAC1 in inflammatory bowel disease: a potential therapeutic target

doi: 10.1016/j.ymthe.2021.06.024

Figure Lengend Snippet: VDAC1 is overexpressed in human colon pathologies and in DSS-treated cells, which result in VDAC1 oligomerization, apoptosis, and mtDNA release that are inhibited by VBIT-12 Immunohistochemical (IHC) staining of VDAC1 was performed on tissue microarray slides obtained from Biomax US (Cat No. CO245). The array contains colon sections from healthy (H, 4 samples), colitis (Co, 4 samples), ulcerative colitis (UC, 4 samples), and Crohn’s disease of the ileocecal junction (CD, 4 samples). (A) Representative sections of the indicated tissues were IHC stained for VDAC1 using specific antibodies. Scale bars are 20 or 50 μm, as indicated. (B) Quantitation of VDAC1 expression levels in the whole area of the provided sections. Staining intensity was quantified using a panoramic microscope and HistoQuant software. Results, mean ± SEM, ∗∗p < 0.01, ∗∗∗p < 0.001, NS not significant (n = 4 for each group). (C–E) Representative images of colon tissue sections from control and DSS-induced colitis mice, IHC, stained for VDAC1 (C; n = 6 for control and n = 7 for DSS). Scale bar is 50 μm. (D–I) CT-26 cells were incubated with the indicated DSS concentration for 48 h in the presence or absence of VBIT-12 (10 or 20 μM) (n = 3). Then samples were analyzed for VDAC1 expression level by immunoblotting with acting serving as loading control (D), oligomerization (E), apoptosis (assayed by PI staining and flow cytometry) (F), intracellular Ca 2+ levels (G), mtROS (H), and mtDNA release (I); all were assayed as described in the Materials and methods section. Results, mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n = 3, NS, not significant. (J) Proposed coupling of VDAC1 overexpression induced by pathological conditions (IBD) and VDAC1 oligomerization mediating the release of apoptogenic proteins from the IMS, leading to apoptosis and the release of mtDNA, leading to inflammasome activation. These processes are inhibited by the VDAC1-interacting molecules, VBIT-4 and VBIT-12, via preventing VDAC1 oligomerization.

Article Snippet: VDAC1 is overexpressed in human colon pathologies and in DSS-treated cells, which result in VDAC1 oligomerization, apoptosis, and mtDNA release that are inhibited by VBIT-12 Immunohistochemical (IHC) staining of VDAC1 was performed on tissue microarray slides obtained from Biomax US (Cat No. CO245).

Techniques: Immunohistochemical staining, Immunohistochemistry, Microarray, Staining, Quantitation Assay, Expressing, Microscopy, Software, Control, Incubation, Concentration Assay, Western Blot, Flow Cytometry, Over Expression, Activation Assay

Journal: bioRxiv

Article Title: Targeting NEDD9-SH3 with a Covalent Peptide Controls Endothelial Phenotype

doi: 10.1101/2025.07.10.663547

Figure Lengend Snippet: a. Paraffin-embedded lung sections from control, rat SU-5416-Hypoxia-Normoxia-PAH, and rat monocrotaline (MCT)-PAH ( n =4-5 rats/condition) or isolated from idiopathic PAH patients ( n =5-8/condition) at the time of lung explant were stained using anti-PECAM1 and anti-NRP-1 antibodies and the immunohistochemical profile in distal pulmonary arterioles was analyzed. Scale bar, 20 μm. b. Human pulmonary artery endothelial cells (HPAECs) and human pulmonary artery smooth muscle cells were co-cultured in a barrier-free system to recapitulate the direct cell-cell contact conditions in PAH lesions. Double immunogold-labeled staining was performed using anti-NRP-1 and anti-Heparan sulfate antibodies, and imaged using transmission electron microscopy. Scale bar, 500 nm. Inset scale bar, 100 nm. c. Disease prevention protocol using the rat SU-5416-hypoxia-normoxia and monocrotaline (MCT) experimental models of PAH and timepoints for intratracheal administration of si-Scrambled (negative) control (Scr) or siRNA against NEDD9. d. The right ventricular systolic pressure (RVSP) measured by cardiac catheterization ( n =3-7 rats/condition) and e. RV mass measured by Fulton Index. IVs, interventricular septum; LV, left ventricle. f. Human pulmonary artery endothelial cells were untreated or treated with VEGF-C (100 nM) for 6 hr and transfected with vehicle (V) control or adenovirus (Ad) containing green fluorescent protein (GFP), Sulf1 cDNA, or siRNA-Sulf1, and cell migration was quantitated using the wound healing assay. Black line, separate experiment iteration without VEGF-C or other treatment ( n =4-7). Blue, DAPI. Scale bar, 400 μm. g. HPAECs were untransfected or transfected with an Ad carrying vehicle (V) control, Scr, Sulf1, or si-Sulf1 and stained with an anti-FAK-Y861 antibody. The number of podosome rosette-positive cells (podosome+) per low power field (40x) and the number of cells with >2 podosomes was quantitated ( n =3). Arrows indicate podosome structures. Scale bar, 50 μm. h. The number of tubes formed per 10x field was counted for untreated HPAECs and cells transfected with Ad carrying V, Sulf1, and si-Sulf1 ( n =3). Scale bar, 300 μm. i . The effect of Ad-Sulf1 on collagen III deposition in HPAECs ( n =5-10). PFE, plaque forming unit (N=1-3 PFEs). Scale bar, 50 μm.

Article Snippet: To analyze the effect of Sulf1 overexpression or inhibition on tube formation, HPAECs were incubated for 24 hours with adenovirus expressing GFP (Ad-GFP), GFP plus human Sulfatase-1 (Ad-Sulf1) or GFP plus human Sulf1-siRNA (Ad-siSulf1), 1 PFU/ml x10 7 (Vector Biolabs) before plating on extracellular matrix-coated slides.

Techniques: Control, Isolation, Staining, Immunohistochemical staining, Cell Culture, Labeling, Transmission Assay, Electron Microscopy, Negative Control, Transfection, Migration, Wound Healing Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: LncRNA FEZF1-AS1 Promotes Multi-Drug Resistance of Gastric Cancer Cells via Upregulating ATG5

doi: 10.3389/fcell.2021.749129

Figure Lengend Snippet: ATG5 knockdown inhibited autophagy of GC cells promoted by overexpression of LncFEZF1-AS1. (A) CCK8 assay on the proliferation of SGC-7901 cells with or without the treatment of FEZF1-AS1, FEZF1-AS1 + ATG5 siRNA or their control. (B) The proliferation of SGC-7901 with or without the treatment of FEZF1-AS1 siRNA, FEZF1-AS1 siRNA + ATG5 or their control group analyzed by CCK8 assay. (C) Representative images of LC3 puncta formation in SGC-7901 observed via IF staining, green: LC3B, bule: DAPI, scar bar: 20 mM. The results represented the mean ± SD of 3 independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were cultured on 24-well chamber slides and co-transfected with FEZF1-AS1 overexpression plasmids and si-ATG5 for 48 h. At the time of harvest, cells were washed with PBS and fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.01% Triton X-100 for 30 min. Then cells were stained with anti-LC3B antibody (Proteintech) overnight at 4°C.

Techniques: Knockdown, Over Expression, CCK-8 Assay, Control, Staining

Journal: Cell Death Discovery

Article Title: hMOF induces cisplatin resistance of ovarian cancer by regulating the stability and expression of MDM2

doi: 10.1038/s41420-023-01478-y

Figure Lengend Snippet: A The volcano plot of transcriptome sequencing in OVCAR3 cells transfected with the hMOF gene and negative control. B Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of differential genes. C Heatmap analysis of significantly altered genes. D The mRNA levels of MDM2, p53, Bcl-2, Bak, Bid, Noxa, and Fas were detected by qRT-PCR in NC cells and hMOF cells. E Western blot analysis of Bax, Bcl-2, p53, MDM2, and hMOF proteins expression before and after treatment with cisplatin (10 µmol/L, 24 h). Compared with NC group, * P < 0.05 and ** P < 0.01 showed significantly difference.

Article Snippet: hMOF-overexpressed OVCAR3 cells were seeded into plates containing glass coverslips and cultured for 24 h. The cell climbing slice was fixed with 4% paraformaldehyde for 2 h, permeabilized in 0.5% PBST (PBS containing 0.5% Triton X-100) for 10 min and blocked with 5% goat serum for 60 min. Then the cell slides were incubated with primary antibodies of hMOF (#ab200660, 1:1000, Abcam) and MDM2 (#66511-1-Ig, 1:500, Proteintech) at 4 °C overnight.

Techniques: Sequencing, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Cell Death Discovery

Article Title: hMOF induces cisplatin resistance of ovarian cancer by regulating the stability and expression of MDM2

doi: 10.1038/s41420-023-01478-y

Figure Lengend Snippet: A Cycloheximide (100 µg/mL) acted on cells at different time points, the degradation rate and half-life of hMOF, MDM2, and p53 proteins were detected by western blot. B Co-immunoprecipitation and immunoblotting for hMOF-MDM2. C Immunofluorescence images for hMOF (green) and MDM2 (red) in hMOF-overexpressed OVCAR3 cells. DAPI was used as a nuclear stain (blue). Colocalization of red and green fluorescence appeared yellow. Double fluorescence assays showing colocalization of hMOF and MDM2 (Pearson’s correlation = 0.669086). D Co-immunoprecipitation and immunoblotting analysis of the acetylation and ubiquitination levels of MDM2 protein. E Western blot was used to detect ubiquitination levels and MDM2 expression in the presence or absence of MG132 (20 µg/mL) in NC cells and hMOF cells. Compared with NC, ** P < 0.01 showed significantly difference.

Article Snippet: hMOF-overexpressed OVCAR3 cells were seeded into plates containing glass coverslips and cultured for 24 h. The cell climbing slice was fixed with 4% paraformaldehyde for 2 h, permeabilized in 0.5% PBST (PBS containing 0.5% Triton X-100) for 10 min and blocked with 5% goat serum for 60 min. Then the cell slides were incubated with primary antibodies of hMOF (#ab200660, 1:1000, Abcam) and MDM2 (#66511-1-Ig, 1:500, Proteintech) at 4 °C overnight.

Techniques: Western Blot, Immunoprecipitation, Immunofluorescence, Staining, Fluorescence, Ubiquitin Proteomics, Expressing

Journal: Cell Death Discovery

Article Title: hMOF induces cisplatin resistance of ovarian cancer by regulating the stability and expression of MDM2

doi: 10.1038/s41420-023-01478-y

Figure Lengend Snippet: A Overexpressed hMOF cells (hMOF cells) were transfected with control shRNA (named hMOF+sh-NC) or shRNA targeting MDM2 (named hMOF+sh-MDM2), and the MDM2 protein expression ( A ) and mRNA level ( B ) were analyzed by western blot and qRT-PCR. C Cells were treated with indicated concentrations of cisplatin for 48 h, cell viability rate was monitored by MTT. D The expressions of Bax, Bcl-2, p53, MDM2 and hMOF were detected by western blot in the presence or absence of cisplatin in cells. The impact for knockdown of MDM2 gene in hMOF cells on apoptosis ( E ) and mitochondrial membrane potential ( F ) were detected by flow cytometry. Compared with NC, * P < 0.05 showed significantly difference. Compared with hMOF+sh-NC, # P < 0.05 and ## P < 0.01 showed significantly difference.

Article Snippet: hMOF-overexpressed OVCAR3 cells were seeded into plates containing glass coverslips and cultured for 24 h. The cell climbing slice was fixed with 4% paraformaldehyde for 2 h, permeabilized in 0.5% PBST (PBS containing 0.5% Triton X-100) for 10 min and blocked with 5% goat serum for 60 min. Then the cell slides were incubated with primary antibodies of hMOF (#ab200660, 1:1000, Abcam) and MDM2 (#66511-1-Ig, 1:500, Proteintech) at 4 °C overnight.

Techniques: Transfection, Control, shRNA, Expressing, Western Blot, Quantitative RT-PCR, Knockdown, Membrane, Flow Cytometry

Journal: Cell Death Discovery

Article Title: hMOF induces cisplatin resistance of ovarian cancer by regulating the stability and expression of MDM2

doi: 10.1038/s41420-023-01478-y

Figure Lengend Snippet: Adenovirus shRNA targeting hMOF was used to knockdown the expression of hMOF in OVCAR3/DDP cells. The tumor growth curve ( A ), tumor weight ( B ), and tumor fluorescence image ( C ) of OVCAR3/DDP cells after being treated with saline or cisplatin in sh-NC and sh-hMOF groups. D Western blot analysis of Bax, Bcl-2, p53, MDM2, and hMOF proteins expression treated with saline or cisplatin in sh-NC and sh-hMOF groups. E TUNEL staining was used to detect the apoptotic cells in sh-NC and sh-hMOF groups treated with saline or cisplatin. Compared with sh-NC+Saline, ** P < 0.01 showed significantly difference. Compared with sh-NC+Cis, # P < 0.05 and ## P < 0.01 showed significantly difference.

Article Snippet: hMOF-overexpressed OVCAR3 cells were seeded into plates containing glass coverslips and cultured for 24 h. The cell climbing slice was fixed with 4% paraformaldehyde for 2 h, permeabilized in 0.5% PBST (PBS containing 0.5% Triton X-100) for 10 min and blocked with 5% goat serum for 60 min. Then the cell slides were incubated with primary antibodies of hMOF (#ab200660, 1:1000, Abcam) and MDM2 (#66511-1-Ig, 1:500, Proteintech) at 4 °C overnight.

Techniques: shRNA, Knockdown, Expressing, Fluorescence, Saline, Western Blot, TUNEL Assay, Staining

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: HHQG alleviates CCl4-induced liver injury in mice. (A) Changes in body weight following CCl 4 -induced liver injury. ( B ) Histopathological changes in liver tissue observed by H&E trichrome staining (200 ×). (C) Histopathological changes in liver tissue observed by Masson’s trichrome staining (200 ×). ( D ) Serum ALT activities. ( E ) Serum AST activities.Data are presented as mean ± SD from three independent experiments.( F ) mRNA expression of IL-6 in liver tissue. ( G ) mRNA expression of TNF-α in liver tissue.( H ) mRNA expression of CCL2 in liver tissue.Data are presented as mean ± SD from three independent experiments. (I) mRNA expression of RASD1 in liver tissue(* p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates no significant difference.)

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Staining, Expressing

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Molecular docking analysis of key active components in HHQG with RASD1. (A) Molecular docking of RASD1 with Quercetin. (B) Molecular docking of RASD1 with Genistein.( C ) Molecular docking of RASD1 with Apigenin.( D ) Molecular docking of RASD1 with Emodin.( E ) Molecular docking of RASD1 with Kaempferol. (In the figures of molecular docking, the stick model represents the active molecule, the red stick structure shows the carbon skeleton of the small-molecule ligands, the green parts are amino acid residues, and the yellow dashed lines indicate chemical bonds formed between active components and amino acid residues.)

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques:

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Optimization of HHQG concentration and construction of RASD1-silenced and overexpressed RAW264.7 cell models for studying inflammatory response regulation. (A) Cell viability of Raw264.7 cells under different concentrations of HHQG. (B) Cell viability of Raw264.7 cells under different concentrations of LPS. ( C ) Cell viability of Raw264.7 cells under different concentrations of HHQG after LPS treatment. (D) Changes in the mRNA expression of RASD1 after its overexpression in Raw264.7 cells. (E) Changes in the mRNA expression of RASD1 at different sites following RASD1 knockdown in Raw264.7 cells. (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group) (Data are presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns indicates no significant difference.)

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Concentration Assay, Expressing, Over Expression, Knockdown

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Effect of RASD1 expression on NLRP3 mRNA levels in LPS-induced RAW264.7 macrophages. (A) Changes in NLRP3 mRNA expression levels following RASD1 overexpression in Raw264.7 cells. (B) Changes in NLRP3 mRNA expression levels at different RASD1 knockdown sites in Raw264.7 cells. (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group) (Data are presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns indicates no significant difference.)

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing, Over Expression, Knockdown

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Effect of RASD1 expression on inflammatory factor mRNA and protein levels in LPS-induced RAW264.7 macrophages. (A) mRNA expression of CCL2 in Raw264.7 cells following RASD1 knockdown and overexpression. (B) mRNA expression of IL-6 in Raw264.7 cells following RASD1 knockdown and overexpression. (C) mRNA expression of TNF-α in Raw264.7 cells following RASD1 knockdown and overexpression. (D) Protein expression of IL-1β in Raw264.7 cell supernatant following RASD1 knockdown and overexpression. (E) Protein expression of IL-6 in Raw264.7 cell supernatant following RASD1 knockdown and overexpression. (F) Protein expression of TNF-α in Raw264.7 cell supernatant following RASD1 knockdown and overexpression. Data are presented as mean ± SD from three independent experiments. (siRASD1 is the RASD1-knockdown group, miRASD1 is the RASD1-overexpression group, * p < 0.05VS.CON; ♆ p < 0.05VS.Model; ◑ p < 0.05VS.HHQG; ▲ p < 0.05VS.Model siRASD1; ● p < 0.05VS.Model miRASD1).

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing, Knockdown, Over Expression

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Effect of RASD1 expression on NLRP3 inflammasome components and downstream inflammatory factors in LPS-induced RAW264.7 macrophages. (A) mRNA expression of NLRP3 in Raw264.7 cells following RASD1 knockdown and overexpression. (B) mRNA expression of ASC in Raw264.7 cells following RASD1 knockdown and overexpression. (C) mRNA expression of caspase-1 in Raw264.7 cells following RASD1 knockdown and overexpression. (D) mRNA expression of GSDMD in Raw264.7 cells following RASD1 knockdown and overexpression. (E) mRNA expression of IL-1β in Raw264.7 cells following RASD1 knockdown and overexpression. (F) mRNA expression of IL-18 in Raw264.7 cells following RASD1 knockdown and overexpression. Data are presented as mean ± SD from three independent experiments. (siRASD1 is the RASD1 knockdown group, miRASD1 represents the RASD1 overexpression group, * p < 0.05VS.CON; ♆ p < 0.05VS.Model; ◑ p < 0.05VS.HHQG; ▲ p < 0.05VS.Model siRASD1; ● p < 0.05VS.Model miRASD1).

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing, Knockdown, Over Expression

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Protein expression of NLRP3, RASD1, and GSDMD in LPS-induced RAW264.7 macrophages following RASD1 modulation. (A) Protein expression of NLRP3, RASD1, and GSDMD in Raw264.7 cells following RASD1 overexpression and the results of the statistical analysis of the strip gray value. (B) Protein expression of NLRP3, RASD1, and GSDMD in Raw264.7 cells following RASD1 knockdown and the results of the statistical analysis of the strip gray value. (Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.) (siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group).

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing, Over Expression, Stripping Membranes, Knockdown

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Effect of RASD1 modulation on PKCδ-NF-κB signaling pathway in RAW264.7 macrophages. (A) mRNA expression of NF-κB genes in Raw264.7 cells following RASD1 knockdown and overexpression. (B) mRNA expression of PKCδ genes in Raw264.7 cells following RASD1 knockdown and overexpression. (C) Protein expression of NF-κB and PKCδ in Raw264.7 cells following RASD1 overexpression and the results of the statistical analysis of the strip gray value. (D) Protein expression of NF-κB and PKCδ in Raw264.7 cells following RASD1 knockdown and the results of the statistical analysis of the strip gray value. (Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.) (Data are presented as mean ± SD from three independent experiments.siRASD1 is the RASD1-knockdown group, and miRASD1 is the RASD1-overexpression group, * p < 0.05VS.CON; ♆ p < 0.05VS.Model; ◑ p < 0.05VS.HHQG; ▲ p < 0.05VS.Model siRASD1; ● p < 0.05VS.Model miRASD1.)

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing, Knockdown, Over Expression, Stripping Membranes

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Effect of HHQG on mRNA expression of NLRP3 inflammasome components and downstream inflammatory factors in CCl4-induced liver injury. (A) mRNA expression of NLRP3 in liver tissue. (B) mRNA expression of ASC in liver tissue. (C) mRNA expression of caspase-1 in liver tissue. (D) mRNA expression of IL-18 in liver tissue. (E) mRNA expression of IL-1β in liver tissue. (F) mRNA expression of GSDMD in liver tissue. (G) Correlation analysis between RASD1 and NLRP3 (r = 0.5727, p < 0.05).

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing

Journal: Scientific Reports

Article Title: HHQG ameliorates acute liver injury (ALI) by inhibiting NLRP3 activation through RASD1-mediated regulation of the PKCδ-NF-κB signaling pathway

doi: 10.1038/s41598-025-24410-z

Figure Lengend Snippet: Effect of HHQG on protein expression of RASD1, NLRP3, and GSDMD in CCl4-induced liver injury. (A) Protein expression of RASD1, GSDMD, and NLRP3 in liver tissue and the results of the statistical analysis of the strip gray value. (B) Immunohistochemical staining of RASD1 in liver tissue (200 ×). (C) Immunohistochemical staining of NLRP3 in liver tissue (200 ×). (D) Immunohistochemical staining of GSDMD in liver tissue (200 ×).(Each band in the figure represents a different blot, all samples are from the same experiment, and the blots were processed in parallel.)

Article Snippet: Carbon tetrachloride (Aladdin, C112044), olive oil (Macklin, O815210), AST/GOT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C010-2–1), ALT/GPT activity kit (Nanjing Jiancheng Bioengineering Institute, China, C009-2–1), paraformaldehyde (Biosharp, BL539A), adhesive slides (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,312–3161), standard coverslips (Jiangsu Shitai Laboratory Equipment Co., Ltd., 80,340–0130), Masson’s trichrome staining kit (Solarbio, G1340), hematoxylin solution (BASO, BA4097), eosin solution (BASO, BA4098), anti-NLRP3 antibody (Bioss, bs-10021r), anti-GSDMD antibody (Abcam, ab219800), anti-RASD1 antibody (Santa Cruz, sc-398988), anti-RASD1 antibody (Proteintech, 12,634–1-AP), anti-IκB-α antibody (Abmart, T55026 ), anti-PKCδ antibody (Abcam, ab182126), anti-β-actin antibody (LABLEAD, A0101-1), goat anti-mouse IgG HRP (LABLEAD, S0100-1), PBS powder (Fuzhou Maixin Biotech Co., Ltd., PBS-0060), antigen retrieval solution (Fuzhou Maixin Biotech Co., Ltd., MVS-0099), citrate buffer for antigen retrieval (Fuzhou Maixin Biotech Co., Ltd., MVS-0101), DAB chromogenic kit (Fuzhou Maixin Biotech Co., Ltd., DAB-0031), ready-to-use immunohistochemistry Elivision Super kit (Fuzhou Maixin Biotech Co., Ltd., KIT-9922), non-immune goat serum (Fuzhou Maixin Biotech Co., Ltd., SP KIT-B3), antibody diluent (Fuzhou Maixin Biotech Co., Ltd., ABD-0030), 3% hydrogen peroxide (Zhongbei Gankong Medical Technology Co., Ltd., 371402ZBGK034), RNAiso Plus (Takara, Japan, 9109), chloroform (Aladdin, C109593), FastKing one-step gDNA removal and cDNA synthesis kit (Tiangen Biotech Co., Ltd., Beijing, China, KR118), FastReal qPCR kit (SYBR Green) (Beijing Tiangen Biotech Co., Ltd., FP217), PVDF membrane (Immobilon®, IPVH00010), fetal bovine serum (WILBER, FBS-SaHu-500), DMEM (Gibco, C11965500BT), LPS (Sigma-Aldrich, L2880), Cell Counting Kit-8 (LABLEAD, CK001), RASD1 overexpression vector and interference knockdown (Suzhou Genepharma Co.,Ltd., D02001), siRNA-mate plus transfection reagent (Suzhou Genepharma Co.,Ltd., G04036 ), Lipofectamine 3000 reagent (LABLEAD, TR002), IL-1β ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02323M1), IL-6 ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02446M1), TNF-α ELISA kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-02415M1).Tribromoethanol for ready use (MeilunBio , MA0478).

Techniques: Expressing, Stripping Membranes, Immunohistochemical staining, Staining