Journal: BMC Oral Health

Article Title: Synergistic proanthocyanidin-copper oxygen-generating microneedle enhances anti-inflammatory activity in dental pulp stem cells and macrophage efferocytosis

doi: 10.1186/s12903-026-07870-1

Figure Lengend Snippet: A : Fluorescence staining of ROS levels in macrophages cultured in the leachate of each hydrogel group; ( B ): Semi-quantitative analysis of ROS fluorescence staining; ( C , D , E) : Expression levels of IL-6, IL-1β, and TNF-α in macrophages cultured in the leachate of each hydrogel group; ( F) : Statistical analysis of macrophage efferocytosis index by immunofluorescence staining; G : Statistical analysis of macrophage efferocytosis index by flow cytometry; ( H ): Immunofluorescence staining to detect macrophage efferocytosis levels (blue: nuclei of macrophages; green: cell membrane of macrophages; red: apoptotic hDPSCs); ( I ): Flow cytometry detection of macrophage efferocytosis levels (FITC-A: F4/80-labeled macrophages; PE-A: CellTracker-labeled apoptotic cells)

Article Snippet: Macrophages (RAW264.7) were incubated for 24 h, and 1 mL of DMEM containing 200 ng/mL LPS was added to induce inflammation for 24 h, then 5 × 10 4 stained apoptotic cells and samples of different groups were added and co-cultured for 24 h. Then, it was fixed with 4% paraformaldehyde for 20 min, washed with PBS, blocked with goat serum for 20 min at room temperature, and incubated with F4/80 antibody (Bosterbio, China) overnight at 4°C.

Techniques: Fluorescence, Staining, Cell Culture, Expressing, Immunofluorescence, Flow Cytometry, Membrane, Labeling

Journal: Cell communication and signaling : CCS

Article Title: HMGB3 promotes the malignant phenotypes and stemness of epithelial ovarian cancer through the MAPK/ERK signaling pathway.

doi: 10.1186/s12964-023-01172-7

Figure Lengend Snippet: Fig. 1 HMGB3 promotes ovarian cancer proliferation. A pLKO.1, HMGB3 shRNA-1 (shHMGB3-1), HMGB3 shRNA-2 (shHMGB3-2), pCMV, and pCMV HMGB3 plasmids were stably transfected into A2780 and SKOV3 cells. HMGB3 protein levels were determined by western blot. B Cells were seeded into 96-well plates and cultured for 1, 2, 3, 4, and 5 days, and an MTT assay was performed to assess cell viability. C A clonogenic assay was used to assess the colony formation efficiency of A2780 and SKOV3 cells with HMGB3 knocked down or overexpressed. D Quantification of the number of clones in C. E Proliferation of A2780 and SKOV3 cells with HMGB3 knocked down or overexpressed detected by EdU assay. Nuclei were stained using DAPI. Scale bar, 10 μm. F Quantification of the ratio of EdU positive cells in (E). Data are presented as the mean ± SEM, *p < 0.05, **p < 0.01, n = 3

Article Snippet: Antibodies against HMGB3 (27465-1-AP), ETS-1 (12118-1-AP), MEK1/2 (11049-1-AP), CCND1 (60186- 1-Ig), c-Myc (10828-1-AP), SOX2 (11064-1-AP), and ALDH1A1 (15910-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: shRNA, Stable Transfection, Transfection, Western Blot, Cell Culture, MTT Assay, Clonogenic Assay, Clone Assay, EdU Assay, Staining

Journal: Cell communication and signaling : CCS

Article Title: HMGB3 promotes the malignant phenotypes and stemness of epithelial ovarian cancer through the MAPK/ERK signaling pathway.

doi: 10.1186/s12964-023-01172-7

Figure Lengend Snippet: Fig. 3 RNA sequencing analysis of signaling pathways involved in HMGB3 function. A2780 cells were transfected with HMGB3 siRNA (siHMGB3) or negative control siRNA (Ctr) for 48 h. High-throughput RNA sequencing analysis was used to compare mRNA expression profiles of the siHMGB3 and Ctr groups. A Volcano plot showing differentially expressed genes (DEGs) between the siHMGB3 and Ctr groups. In total, 91 genes were up-regulated (Up) and 679 genes were down-regulated (Down); other genes expression levels were not significantly altered (no-DEGs) (B) Kyoto Encyclopedia of Genes and Genomes enrichment analysis of genes down-regulated in the siHMGB3 group relative to the Ctr group. C Heatmap showing genes involved in stem cell pluripotency and the MAPK signaling pathway down-regulated in the siHMGB3 group relative to the Ctr group. D A2780 cells were transfected with HMGB3 siRNA (siHMGB3) or negative control siRNA (Ctr) for 48 h, and qRT-PCR used to verify down-regulation of representative genes in the siHMGB3 group relative to the Ctr group. Data are presented as the mean ± SEM, *p < 0.05, **p < 0.01, n = 3

Article Snippet: Antibodies against HMGB3 (27465-1-AP), ETS-1 (12118-1-AP), MEK1/2 (11049-1-AP), CCND1 (60186- 1-Ig), c-Myc (10828-1-AP), SOX2 (11064-1-AP), and ALDH1A1 (15910-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: RNA Sequencing, Protein-Protein interactions, Transfection, Negative Control, High Throughput Screening Assay, Expressing, Quantitative RT-PCR

Journal: Cell communication and signaling : CCS

Article Title: HMGB3 promotes the malignant phenotypes and stemness of epithelial ovarian cancer through the MAPK/ERK signaling pathway.

doi: 10.1186/s12964-023-01172-7

Figure Lengend Snippet: Fig. 4 HMGB3 activates the MAPK/ERK signaling pathway in ovarian cancer cells. A p-MEK1/2, MEK1/2, p-ERK1/2, ERK1/2, ETS-1, CCND1, c-Myc, HMGB3, and β-actin protein levels of in A2780 and SKOV3 cells with HMGB3 knocked down or overexpressed detected by western blot. B Quantification of the protein levels in (A). C Protein levels in ovarian cancer cells transfected with pLKO.1, HMGB3 shRNA-1 (shHMGB3-1) and/or pCMV HMGB3 detected by western blot. (D) Quantification of the protein levels in (C). Data are presented as the mean ± SEM, *p < 0.05, **p < 0.01, n = 3

Article Snippet: Antibodies against HMGB3 (27465-1-AP), ETS-1 (12118-1-AP), MEK1/2 (11049-1-AP), CCND1 (60186- 1-Ig), c-Myc (10828-1-AP), SOX2 (11064-1-AP), and ALDH1A1 (15910-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Western Blot, Transfection, shRNA

Journal: Cell communication and signaling : CCS

Article Title: HMGB3 promotes the malignant phenotypes and stemness of epithelial ovarian cancer through the MAPK/ERK signaling pathway.

doi: 10.1186/s12964-023-01172-7

Figure Lengend Snippet: Fig. 6 HMGB3 promotes the malignant phenotypes of ovarian cancer via the MAPK/ERK signaling pathway. A A2780 and SKOV3 cells with HMGB3 overexpression were seeded into 6 cm dishes and then treated with or without AZD6244 (5 µM)/PD0325901 (10 µM) for 24 h. p-ERK1/2, ERK1/2, HMGB3, and β-actin protein levels detected by western blot. B A2780 and SKOV3 cells with HMGB3 overexpression were seeded into 96-well plates and then treated with or without AZD6244 (5 µM)/PD0325901 (10 µM) for 72 h. Cell viability was detected by MTT assay. C A2780 and SKOV3 cells with HMGB3 overexpression were treated with or without AZD6244 (1 µM)/PD0325901 (2 µM) for 1–2 weeks. Colony formation efficiency was assessed by clonogenic assay. D Quantification of the number of clones in (C). E A2780 and SKOV3 cells with HMGB3 overexpression were seeded into transwell plates and then treated with or without AZD6244 (5 µM)/PD0325901 (10 µM) for 12–24 h. Cell migration and invasion were evaluated by transwell assay. Scale bar, 50 μm. F Quantification of the number of cells in (E). Data are presented as the mean ± SEM, **p < 0.01, n = 3

Article Snippet: Antibodies against HMGB3 (27465-1-AP), ETS-1 (12118-1-AP), MEK1/2 (11049-1-AP), CCND1 (60186- 1-Ig), c-Myc (10828-1-AP), SOX2 (11064-1-AP), and ALDH1A1 (15910-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Over Expression, Western Blot, MTT Assay, Clonogenic Assay, Clone Assay, Migration, Transwell Assay

Journal: Cell communication and signaling : CCS

Article Title: HMGB3 promotes the malignant phenotypes and stemness of epithelial ovarian cancer through the MAPK/ERK signaling pathway.

doi: 10.1186/s12964-023-01172-7

Figure Lengend Snippet: Fig. 7 HMGB3 promotes ovarian cancer cell proliferation in vivo through the MAPK/ERK signaling pathway. A2780 cells (5 × 106) transfected with pLKO.1, HMGB3 shRNA-1 (shHMGB3-1), or HMGB3 shRNA-2 (shHMGB3-2) were subcutaneously injected into nude mice. Mice were divided into three groups: pLKO.1 (Ctr), shHMGB3-1, and shHMGB3-2 (n = 6 per group). Two weeks post-injection, mice were euthanized and the xenograft tumors were removed. A Tumors from each group are shown. B Tumor volumes of each group. C Body weight of each group. D p-MEK1/2, MEK1/2, p-ERK1/2, ERK1/2, HMGB3, and β-actin protein levels in tumor tissues detected by western blot. E Quantification of the protein levels in (D). A2780 cells (5 × 106) transfected with pCMV or pCMV HMGB3 were subcutaneously injected into nude mice. Mice were divided into three groups: pCMV (Ctr), pCMV HMGB3, and pCMV HMGB3 + AZD6244 (n = 6 per group). One group of mice received an intraperitoneal injection of AZD6244 (25 mg/ kg) once a day. Two weeks post-injection, mice were euthanized and xenograft tumors were removed. F Tumors from each group are shown. G Tumor volumes of each group. H Body weight of each group. I p-MEK1/2, MEK1/2, p-ERK1/2, ERK1/2, HMGB3, and β-actin protein levels in tumor tissues detected by western blot. J Quantification of the protein levels in (I). Data are presented as mean ± SEM, #p > 0.05, **p < 0.01, n = 6

Article Snippet: Antibodies against HMGB3 (27465-1-AP), ETS-1 (12118-1-AP), MEK1/2 (11049-1-AP), CCND1 (60186- 1-Ig), c-Myc (10828-1-AP), SOX2 (11064-1-AP), and ALDH1A1 (15910-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: In Vivo, Transfection, shRNA, Injection, Western Blot

Journal: Cell communication and signaling : CCS

Article Title: HMGB3 promotes the malignant phenotypes and stemness of epithelial ovarian cancer through the MAPK/ERK signaling pathway.

doi: 10.1186/s12964-023-01172-7

Figure Lengend Snippet: Fig. 8 Schematic summary of the study findings. HMGB3 overexpression activates the MAPK/ERK signaling pathway, thereby promoting ovarian cancer proliferation, metastasis, and stemness. Inhibition of the MAPK/ERK signaling pathway using specific inhibitors counteracts the effects of HMGB3

Article Snippet: Antibodies against HMGB3 (27465-1-AP), ETS-1 (12118-1-AP), MEK1/2 (11049-1-AP), CCND1 (60186- 1-Ig), c-Myc (10828-1-AP), SOX2 (11064-1-AP), and ALDH1A1 (15910-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Over Expression, Inhibition

Journal: Cell death & disease

Article Title: High glucose levels promote glycolysis and cholesterol synthesis via ERRα and suppress the autophagy-lysosomal pathway in endometrial cancer.

doi: 10.1038/s41419-025-07499-y

Figure Lengend Snippet: Fig. 3 After stimulation by high glucose, the role of ERRα in regulating glucose and cholesterol metabolism. A Heatmap of the average read density and chromatin occupancy of the transcription start site (TSS) of ERRα in two independent samples of HEC-1A cells with control or upregulated ERRα. B, C Distribution of CUT&Tag peaks of ERRα in the genomic region from the second repetition of control samples. D, E Distribution of ERRα sites relative to HK2 and HMGCS1 promoters based on the CUT&Tag analysis. F HEC-1A and KLE EC cells transduced with si-ERRα were treated with NG or HG for 48 h. The expression of ERRα, HK2, PKM2, HMGCR, and HMGCS1 was measured by western blotting (WB), with β-actin used as a loading control. G HEC-1A and KLE EC cells with ov-ERRα were subjected to NG or HG for 48 h. The expression of ERRα, HK2, PKM2, HMGCR, and HMGCS1 was measured by western blotting, with β-actin used as a loading control. H, I Analysis of the data in panel F comparing the differences in protein expression among the si-ERRα-NG, con-HG, and si-ERRα-HG groups using the con- NG group as the reference baseline. J, K Analysis of the data in panel G comparing the differences in protein expression among the con-NG, ov-ERRα-HG, and con-HG groups using the ov-ERRα-NG group as the reference baseline. L–O HEC-1A and KLE EC cells were treated with Mnt, NG, or HG for 48 h, after which the lactate or TC concentrations were measured with three replications, respectively. P, S HEC-1A and KLE EC cells with or without ERRα knockdown were treated with NG or HG for 48 h, after which the lactate or TC concentration was measured with three replications, respectively. *P < 0.05, **P < 0.01 or ***P < 0.001. NG normal glucose, HG high glucose, EC endometrial cancer, TC total cholesterol.

Article Snippet: Membranes were incubated with antibodies specific for ERRα (1:500, #ab137489, Abcam, MA, USA), HK2 (1:1000, #66974- 1-Ig, Proteintech, Wuhan, China), HK2, (#ab209847, Abcam, MA, USA), PKM2 (1:1000, #60268-1-Ig, Proteintech, Wuhan, China), HMGCS1 (1:1000, #17643-1- AP, Proteintech, Wuhan, China), HMGCR (1:1000, #A16875, ABclonal, Wuhan, China), p62 (1:1000, #ab207305, Abcam, MA, USA) and β-actin (1:1000, #YM8343, ImmunoWay, Suzhou, China); washed; and incubated with a goat anti-rabbit secondary antibody (1:10,000, #SA00001-2, Proteintech, Wuhan, China) or goat anti-mouse secondary antibody (1:10,000, #SA00001-1, Proteintech, Wuhan, China) for 2 h. The membranes were washed 30min.

Techniques: Control, Transduction, Expressing, Western Blot, Knockdown, Concentration Assay

Journal: Cancer Research

Article Title: MEN1 Promotes Ferroptosis by Disrupting CD44 Alternative Splicing to Suppress Lung Cancer

doi: 10.1158/0008-5472.CAN-25-0021

Figure Lengend Snippet: MEN1 -modulated ferroptosis is required for suppressing lung tumor growth and metastasis. A–D, Cell viability and representative invasion and migration images of WT and KO NCI-H460 cells; invaded cells and migration rate were quantified at the indicated time points. E, Schematic of experimental lung metastasis model in M-NSG mice. F, Representative IHC (Ki67 and HMGB1) and H&E staining images of lung tissue sections from M-NSG mice xenografted with WT/KO NCI-H460 cells. Scale bars, 1 mm or 50 μm. G and H, Tumor burden ( G ) and quantification of Ki67/HMGB1 IHC staining in lung tissues ( H ) from xenografted M-NSG mice. I, Schematic of the primary mouse lung cancer study ( n = 7, 9, and 8 for WT, KS, and KMS cohorts). J–M, Representative lung pictures ( J ), H&E staining and Ki67/HMGB1 IHC images of lung tissues ( K ), tumor burden and tumor number ( L ), and quantification of Ki67/HMGB1 IHC staining ( M ). Scale bars, 1 mm or 50 μm. Black arrows, tumor nodules. N, Kaplan–Meier survival analysis of WT, KS, and KMS mice. O, Lipid peroxidation (LPO) levels in lungs from WT, KS, and KMS mice. a.u, arbitrary unit. P and Q, Representative RNA FISH images ( P ) and quantification of GPX4 probe numbers in WT lung tissues and KS/KMS lung tumor tissues ( Q ). Scale bars, 20 or 50 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: The paraffin-embedded tissue sections after deparaffinization and rehydration were subjected to antigen retrieval in 0.1 mol/L citrate buffer (pH 6.0) in a steam pot for 15 minutes, blocked with 5% BSA for 1 hour, and incubated with primary antibodies including menin (Bethyl Laboratories, #A300-105A, RRID: AB_2143306), Ki67 (Cell Signaling Technology, #9129, RRID: AB_2687446), HMGB1 (Proteintech, #10829-1-AP, RRID: AB_2232989), and SFTPC (Abclonal, #A1835, RRID: AB_2763872).

Techniques: Migration, Staining, Immunohistochemistry

Journal: Scientific Reports

Article Title: TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis

doi: 10.1038/srep24859

Figure Lengend Snippet: MIN6 cells were plated at 5 × 10 5 cells per well in 6-well plates and exposed to a TCF7L2-specific short hairpin RNA (shTCF7L2) or a scrambled shRNA (shScr) for 72 h, then cultured for 24 h in the presence of 100 nM E 2 or 1 μM P 4 . ( A ) Western blots showing the TCF7L2 protein content after E 2 or P 4 treatment. ( B ) Viable cells. ( C,D ) Basal and stimulated insulin/proinsulin secretions (normalized to viable cell numbers). ( E,F ) Stimulatory indexes. ( G ) Proinsulin-to-insulin ratio. * P < 0.05 vs. sex hormone treatment control; # P < 0.05 shTCF7L2 vs. shScr.

Article Snippet: The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA).

Techniques: shRNA, Cell Culture, Western Blot, Control

Journal: Scientific Reports

Article Title: TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis

doi: 10.1038/srep24859

Figure Lengend Snippet: MIN6 cells were plated at 5 × 10 5 cells per well in 6-well plates and exposed to high glucose concentration (33.3 mM) and transfected with TCF7L2-IRES2-EGFP (OE-TCF7L2) or a control vector (CV) for 72 h, then cultured for 24 h in the presence of 100 nM E 2 or 1 μM P 4 . ( A ) Western blots showing the TCF7L2 protein content after E 2 or P 4 treatment. ( B ) Viable cells. ( C , D ) Basal and stimulated insulin/proinsulin secretions (normalized to viable cell numbers). ( E,F ) Stimulatory indexes. ( G ) Proinsulin-to-insulin ratio. * P < 0.05 vs. sex hormone treatment control; # P < 0.05 OE-TCF7L2 vs. CV.

Article Snippet: The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA).

Techniques: Concentration Assay, Transfection, Control, Plasmid Preparation, Cell Culture, Western Blot

Journal: Scientific Reports

Article Title: TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis

doi: 10.1038/srep24859

Figure Lengend Snippet: HepG2 cells (2.5 × 10 5 cells per well) were seeded in 6-well plates and exposed to a TCF7L2-specific short hairpin RNA (shTCF7L2) or a scrambled shRNA (shScr) for 72 h, or transfected with TCF7L2-IRES2-EGFP (OE-TCF7L2) or a control vector (CV) for 72 h, then cultured for 24 h in the presence of 100 nM E 2 or 1 μM P 4 . ( A,B ) 2-NBDG uptake. ( C,D ) Glucose production. ( E,F ) Western blot. * P < 0.05 vs. sex hormone treatment control; # P < 0.05 shTCF7L2 vs. shScr or OE-TCF7L2 vs. CV.

Article Snippet: The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA).

Techniques: shRNA, Transfection, Control, Plasmid Preparation, Cell Culture, Western Blot

Journal: Experimental and Therapeutic Medicine

Article Title: Increased susceptibility to ischemia-induced ventricular tachyarrhythmias in depressed rats: Involvement of reduction of connexin 43

doi: 10.3892/etm.2011.396

Figure Lengend Snippet: Representative immunoblots of the Cx43 protein. Quantitative densitometric analysis of the amount of NP-Cx43 and T-Cx43 normalized to GAPDH (NP-Cx43/GAPDH; T-Cx43/GAPDH). The relative amounts of NP-Cx43 and T-Cx43 were expressed as a percentage of the control-SO or CMS-SO groups. (A) Comparison of the amount of total Cx43 between the control-SO and CMS-SO groups. #P<0.05 vs. control-SO group. (B) Comparison of the amount of total Cx43 and NP-Cx43 between the control-SO and CMS-MI groups in CMS rats. #P<0.05 vs. CMI-SO group. T-Cx43 = NP-Cx43 + P-Cx43. NP-Cx43, non-phosphorylated Cx43; P-Cx43, phosphorylated Cx43; T-Cx43, total Cx43; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; SO, sham-operated; MI, myocardial ischemia; CMS, chronic mild stress; Cx43, connexin 43.

Article Snippet: Immunoblot analysis Regions of pulverized frozen ischemia from left ventricle samples were analyzed by quantitative immunoblotting using a goat polyclonal anti-Cx43 antibody (sc-56698; Santa Cruz Biotechnology, CA, USA) for total Cx43 protein and a non-phosphorylated mouse monoclonal anti-Cx43 antibody (13–8300; Zymed/Invitrogen, Carlsbad, CA, USA) for non-phosphorylated Cx43 content.

Techniques: Western Blot, Control, Comparison

Journal: Experimental and Therapeutic Medicine

Article Title: Increased susceptibility to ischemia-induced ventricular tachyarrhythmias in depressed rats: Involvement of reduction of connexin 43

doi: 10.3892/etm.2011.396

Figure Lengend Snippet: Representative immunofluorescence images of the Cx43 protein. SO, sham-operated; MI, myocardial ischemia; CMS, chronic mild stress. #P<0.05 vs. control-SO group. Bars, 100 μm. Cx43, connexin 43.

Article Snippet: Immunoblot analysis Regions of pulverized frozen ischemia from left ventricle samples were analyzed by quantitative immunoblotting using a goat polyclonal anti-Cx43 antibody (sc-56698; Santa Cruz Biotechnology, CA, USA) for total Cx43 protein and a non-phosphorylated mouse monoclonal anti-Cx43 antibody (13–8300; Zymed/Invitrogen, Carlsbad, CA, USA) for non-phosphorylated Cx43 content.

Techniques: Immunofluorescence, Control

Journal: Nature Communications

Article Title: Cholecystectomy-related gut microbiota dysbiosis exacerbates colorectal tumorigenesis

doi: 10.1038/s41467-025-62956-8

Figure Lengend Snippet: a Volcano plot of RNA-seq analysis of colon cancerous tissue. The upregulated genes (green) and the downregulated genes (red) with a fold change <0.6 ( n = 3/group) were shown. P values were adjusted by Benjamini & Hochberg (BH) method to control FDR. FDR-adjusted P < 0.05 was shown. b GSEA of the changes in the FXR pathway genes between Sham+AOM/DSS and GBx+AOM/DSS group ( n = 3/group). Negative NES indicates that the level is lower in GBx+AOM/DSS group. c Relative expression of FXR in colon cancerous tissues ( n = 7/group). d Relative expression of SHP in colon cancerous tissues ( n = 6 vs 5). e KEGG pathway analysis of Sham+AOM/DSS and GBx+AOM/DSS mice ( n = 3/group). P values were adjusted by BH method to control FDR. FDR-adjusted P < 0.05 was shown. f Relative expression of MYC in colon cancerous tissues ( n = 7/group). g Western blot analysis of FXR, β-catenin, c-Myc, and GAPDH within the colon cancerous tissues from Sham+AOM/DSS and GBx+AOM/DSS mice. GAPDH was used as a loading control. h Immunofluorescence analysis of β-catenin ( n = 7/group). β-Catenin, green; DAPI, blue. Scale bar, 20 μm. i Representative image of IHC staining of FXR, β-catenin, and c-Myc ( n = 7/group). The bottom scale bar is 200 μm and the top scale bar is 20 μm. j Co-immunoprecipitation analysis of the interaction between FXR and β-catenin in colon cancerous tissues. k Co-IP of the interaction between TCF4 and β-catenin in colon cancerous tissues. Data are shown as the mean ± SEM. P values were determined by two-tailed t test and were indicated in each figure. FXR farnesoid X receptor, SHP small heterodimer partner, TCF transcription factor. All data are representative of more than three independent experiments. Source data are provided as a Source Data file.

Article Snippet: Azoxymethane (AOM) and dextran sodium sulfate (DSS) were purchased from MP Biomedicals (Shanghai) Co., Ltd. Antibodies used were as follows: FXR monoclonal antibody (Santa cruz, sc-25309), β-catenin monoclonal antibody (Proteintech, 66379-1-Ig), c-Myc monoclonal antibody (Abcam, ab32072), Ki67 monoclonal antibody (Cell Signaling Technology, 9449), TCF4 monoclonal antibody (MCE, HY- P80520 ), and GAPDH polyclonal antibody (Proteintech, 10494-1-AP).

Techniques: RNA Sequencing, Control, Expressing, Western Blot, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Co-Immunoprecipitation Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Cholecystectomy-related gut microbiota dysbiosis exacerbates colorectal tumorigenesis

doi: 10.1038/s41467-025-62956-8

Figure Lengend Snippet: a Schematic overview shows OCA treatments in the sham or cholecystectomy mice with AOM/DSS treatment. b Representative images of the colon ( n = 6/group). c Average tumor number ( n = 6/group). d Representative images of H&E staining (left) and semiquantitative analysis (right) ( n = 6/group). H&E staining showed normal, dysplastic mucosae and carcinoma in the colon tissues. The bottom scale bar is 200 μm and the top scale bar is 20 μm. e Protein expression of FXR, β-catenin, c-Myc, and GAPDH within the colon cancerous tissues for each group. f Co-immunoprecipitation of the interaction between FXR and β-catenin in colon cancerous tissues. g Co-immunoprecipitation of the interaction between TCF4 and β-catenin in colon cancerous tissues. h Representative image and quantification of organoids generated from GBx+AOM/DSS mice treated with control, GUDCA (100 μM), or GUDCA + OCA (10 μM) ( n = 5/group). Scale bar, 20 μm. i IHC staining of FXR, β-catenin, and c-Myc in healthy controls and cholecystectomy patients in colon tissues. ( n = 4/group). The bottom scale bar is 200 μm and the top scale bar is 20 μm. Data are shown as the mean ± SEM. P values were determined by ANOVA and were indicated in each figure. HC healthy control, GBx cholecystectomy, OCA obeticholic acid. Source data are provided as a Source Data file.

Article Snippet: Azoxymethane (AOM) and dextran sodium sulfate (DSS) were purchased from MP Biomedicals (Shanghai) Co., Ltd. Antibodies used were as follows: FXR monoclonal antibody (Santa cruz, sc-25309), β-catenin monoclonal antibody (Proteintech, 66379-1-Ig), c-Myc monoclonal antibody (Abcam, ab32072), Ki67 monoclonal antibody (Cell Signaling Technology, 9449), TCF4 monoclonal antibody (MCE, HY- P80520 ), and GAPDH polyclonal antibody (Proteintech, 10494-1-AP).

Techniques: Staining, Expressing, Immunoprecipitation, Generated, Control, Immunohistochemistry

Journal: Nature Communications

Article Title: Cholecystectomy-related gut microbiota dysbiosis exacerbates colorectal tumorigenesis

doi: 10.1038/s41467-025-62956-8

Figure Lengend Snippet: Cholecystectomy can induce gut microbiota disorder, characterized by a significant decrease in the abundance of B. breve and an increase in the abundance of R. gnavus . The reduction in B. breve elevates the level of TUDCA via its BSH activity. Meanwhile, the augmented R. gnavus can promote the formation of TUDCA through its 7β-HSDH activity. TUDCA may ultimately lead to colon carcinoma by disrupting the interaction between FXR and β-catenin, which could further increase the binding of β-catenin and TCF4, thereby promoting MYC expression and facilitating colorectal tumorigenesis. OCA treatment may ultimately prevent colon carcinoma through FXR activation. The schematic was created using Adobe Illustrator 2024 (v28.7.0). B. breve , Bifidobacterium breve , R. gnavus, Ruminococcus gnavus ; TUDCA tauroursodeoxycholic acid, 7β-HSDH 7β-hydroxysteroid dehydrogenase, BSH bile salt hydrolase, FXR farnesoid X receptor, TCF4 transcription factor 4, OCA obeticholic acid.

Article Snippet: Azoxymethane (AOM) and dextran sodium sulfate (DSS) were purchased from MP Biomedicals (Shanghai) Co., Ltd. Antibodies used were as follows: FXR monoclonal antibody (Santa cruz, sc-25309), β-catenin monoclonal antibody (Proteintech, 66379-1-Ig), c-Myc monoclonal antibody (Abcam, ab32072), Ki67 monoclonal antibody (Cell Signaling Technology, 9449), TCF4 monoclonal antibody (MCE, HY- P80520 ), and GAPDH polyclonal antibody (Proteintech, 10494-1-AP).

Techniques: Activity Assay, Binding Assay, Expressing, Activation Assay