oct4 octamer binding transcription factor 4  (ReproCELL)

Journal: Stem Cells Translational Medicine

Article Title: Microencapsulated 3D culture of human umbilical cord-derived mesenchymal stem cells enhances their therapeutic effect on intrauterine adhesion

doi: 10.1093/stcltm/szaf044

Figure Lengend Snippet: The cell proliferation and stemness of 3D-hUCMSCs. (A) Microscopic morphology of 3D-hUCMSCs. Scale bars: 100 µm. (B) Statistical analysis of cell counts on day 1, 3, and 5 of 3D-hUCMSCs. At least 6 representative visual fields for each were counted. (C) Proliferation of hUCMSCs detected by CCK8 ( n = 3). (D) Western blot results of OCT4, SOX2, KLF4, and c-MYC. (E-H) Relative mRNA expression of OCT4, SOX2, KLF4, and c-MYC adjusted to GAPDH ( n = 3). 3D-hUCMSCs, 3D culture of human umbilical cord-derived mesenchymal stem cells.

Article Snippet: PVDF membranes were incubated overnight at 4 °C with primary antibodies against octamer-binding transcription factor-4 (OCT4) (CST, 1:2000, US), SRY-related high-mobility-group-box protein-2 (SOX2) (CST, 1:2000, US), Kruppel-like factor-4 (KLF4) (Abcam, 1:2000, UK), cellular-myelocytomatosis viral oncogene (c-MYC) (Abcam, 1:2000, UK), TGF-β1 (Abcam, 1:2000, UK), Smad3 (Abcam, 1:2000, UK), and pSmad3 (Abcam, 1:2000, UK) after blocking with 5% skim milk.

Techniques: Western Blot, Expressing, Derivative Assay

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Huaier enhances the tumor-killing effect and reverses gemcitabine-induced stemness by suppressing FoxM1.

doi: 10.1016/j.phymed.2024.155656

Figure Lengend Snippet: Fig. 2. Huaier inhibits pancreatic cancer stemness. (A-C) Sphere formation assays proved that Huaier reduced the PC stemness by reducing the diameter of sphere colonies. Scale bar = 200 µm. (D) Flow cytometry demonstrated that Huaier (90 µg/ml) treatment decreased the CD44-positive cell number. (E-H) Western blotting experiments confirmed that OCT4, CD44, and SOX2 expression significantly decreased with increasing Huaier concentration. NC: negative control, *: Compared to the control group, p < 0.05.

Article Snippet: Primary antibodies against β-actin (No. 81115-1-RR), FoxM1 (No. 13147-1-AP), CD44 (No. 13147-1-AP), octamer-binding transcription Factor 4 (OCT4) (No. 11263-1-AP), and SRY-box transcription Factor 2 (SOX2) (No. 11064-1-AP) and secondary antibodies were obtained from Proteintech (Rosemont, IL, USA).

Techniques: Flow Cytometry, Western Blot, Expressing, Concentration Assay, Negative Control, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Huaier enhances the tumor-killing effect and reverses gemcitabine-induced stemness by suppressing FoxM1.

doi: 10.1016/j.phymed.2024.155656

Figure Lengend Snippet: Fig. 5. Huaier reverses gemcitabine-induced cancer stemness in vitro. (A-C) The sphere-forming ability of PC cells after gemcitabine (1 µM), Huaier (90 µg/ml) or gemcitabine plus Huaier treatment. The diameter of stem spheres was measured for statistical analysis. Scale bar = 200 µm. (D-G) Western blotting detected the expression of CD44, SOX2, and OCT4 after treatment with gemcitabine (1 µM), Huaier (90 µg/ml), or gemcitabine plus Huaier. GEM: gemcitabine. *: Compared to the control group, p < 0.05. Scale bar = 200 µm.

Article Snippet: Primary antibodies against β-actin (No. 81115-1-RR), FoxM1 (No. 13147-1-AP), CD44 (No. 13147-1-AP), octamer-binding transcription Factor 4 (OCT4) (No. 11263-1-AP), and SRY-box transcription Factor 2 (SOX2) (No. 11064-1-AP) and secondary antibodies were obtained from Proteintech (Rosemont, IL, USA).

Techniques: In Vitro, Western Blot, Expressing, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Huaier enhances the tumor-killing effect and reverses gemcitabine-induced stemness by suppressing FoxM1.

doi: 10.1016/j.phymed.2024.155656

Figure Lengend Snippet: Fig. 6. The effect of Huaier on reducing pancreatic cancer stem cell-like properties is FoxM1-dependent (A-B) CCK8 assays demonstrated that thiostrepton treatment reduced the cell viability significantly, and the thiostrepton (1 µM) plus gemcitabine (50 nM) group showed the lowest cell viability. (C-D) Western blotting showed that after thiostrepton (1 µM) or gemcitabine (1 µM) treatment, CDK6 and cyclinD1 expression was suppressed, and the combined group showed the lowest expression. (E-F) Colony formation assays showed that thiostrepton (1 µM) plus gemcitabine (1 µM) showed the most effective tumor proliferation suppression effect compared with either drug alone. (G-I) Western blotting detected CD44, SOX2, and OCT4 in the thiostrepton (1 µM), gemcitabine (1 µM), and thiostrepton plus gemcitabine groups. (J-L) Huaier (90 µg/ml) no longer reduced FoxM1 and CSC-associated proteins after thiostrepton (1 µM) treatment. Th: Thiostrepton *: Compared to the control group, p < 0.05.

Article Snippet: Primary antibodies against β-actin (No. 81115-1-RR), FoxM1 (No. 13147-1-AP), CD44 (No. 13147-1-AP), octamer-binding transcription Factor 4 (OCT4) (No. 11263-1-AP), and SRY-box transcription Factor 2 (SOX2) (No. 11064-1-AP) and secondary antibodies were obtained from Proteintech (Rosemont, IL, USA).

Techniques: Western Blot, Expressing, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Huaier enhances the tumor-killing effect and reverses gemcitabine-induced stemness by suppressing FoxM1.

doi: 10.1016/j.phymed.2024.155656

Figure Lengend Snippet: Fig. 7. Huaier enhanced the antitumor effect of gemcitabine by inhibiting FoxM1 and reversing cancer stemness in vivo. (A) Huaier (50 mg/kg and 100 mg/kg) reduced the progression of pancreatic cancer compared with the NC group. Thiostrepton (17 mg/kg) was used as a positive control. (B) Pattern diagram of a subcutaneous tumor model in nude mice. (C) The statistical chart of tumor size and weight after Huaier and thiostrepton intervention. (D-E) The tumor size and weight after gemcitabine (50 mg/kg), Huaier (100 mg/kg) or gemcitabine plus Huaier treatment. (F-G) Immunohistochemicals detected the expression of FoxM1, CD44, OCT4, and SOX2 in vivo after gemcitabine, Huaier or gemcitabine plus Huaier treatment. Scale bar = 200 µm. GEM: gemcitabine. *: Compared to the control group, p < 0.05.

Article Snippet: Primary antibodies against β-actin (No. 81115-1-RR), FoxM1 (No. 13147-1-AP), CD44 (No. 13147-1-AP), octamer-binding transcription Factor 4 (OCT4) (No. 11263-1-AP), and SRY-box transcription Factor 2 (SOX2) (No. 11064-1-AP) and secondary antibodies were obtained from Proteintech (Rosemont, IL, USA).

Techniques: In Vivo, Positive Control, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: PIM Kinase Inhibition Attenuates the Malignant Progression of Metastatic Hepatoblastoma

doi: 10.3390/ijms25010427

Figure Lengend Snippet: PIM kinase inhibition decreased metastatic hepatoblastoma cell stemness markers. ( A ) HLM_2 (1 × 10 6 ) cells were treated with increasing concentrations of AZD1208 (0, 10, or 20 µM) for 72 h. The RNA was collected and qPCR completed. Treatment with AZD1208 (10 µM) decreased the mRNA abundance of each of the markers; Nanog (1.00 ± 0.0 v. 0.45 ± 0.47, control v. AZD1208 10 µM, p ≤ 0.05), SOX2 (1.00 ± 0.0 v. 0.25 ± 0.34, control v. AZD1208 10 µM, p ≤ 0.001), Nestin (1.00 ± 0.0 v. 0.69 ± 0.11, control v. AZD1208 10 µM, p ≤ 0.001), and OCT4 (1.00 ± 0.0 v. 0.39 ± 0.39, control v. AZD1208 10 µM, p ≤ 0.001). Similar results were seen following treatment with 20 µM concentration of AZD1208. ( B ) Immunoblotting demonstrated a decrease in protein expression of stemness markers Nestin, Oct4, and Nanog with increase concentrations of AZD1208 (0–20 µM). Vinculin served as a loading control. Data are reported as mean ± SEM and represent at least three biologic replicates. * p ≤ 0.05, *** p ≤ 0.001.

Article Snippet: Probes for NESTIN , octamer-binding transcription factor 4 ( OCT4 ), homeobox protein NANOG , sex determining region Y-box 2 ( SOX2 ), and β-ACTIN were obtained (Applied Biosystems, Foster City, CA, USA) and checked for non-specific binding using the basic local alignment as previously described [ , ] qPCR was performed with 10 ng cDNA in 20 μL reaction volume.

Techniques: Inhibition, Concentration Assay, Western Blot, Expressing

Journal: BMC cancer

Article Title: Wnt/β-catenin pathway is a key signaling pathway to trastuzumab resistance in gastric cancer cells.

doi: 10.1186/s12885-023-11447-4

Figure Lengend Snippet: Fig. 1 Stemness and EMT-like phenotypes of trastuzumab-resistant gastric cancer. (a) NCI-N87R cells had significantly larger sphere size and comprised higher cell counts of spheres larger than 50 μm compared with NCI-N87 cells. (b) E-cadherin was significantly downregulated in NCI-N87R cells, as confirmed by immunofluorescence, compared with parental cells. Stem cell markers including CD44s, CD54, BMI1, Oct4, Vimentin and Snail were significantly upregulated in NCI-N87R cells compared with parental cells, according to Western blot analysis. *P < 0.05, **P < 0.01, ***P < 0.005

Article Snippet: Next, the membrane was incubated overnight at 4 °C with rabbit anti-polycomb complex protein BMI-1 (BMI1) (1: 500; #ab135713; Abcam, Cambridge, UK), rabbit anti-HER2 (1: 1,000; #2165; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Snail (1: 500; #3895; Cell Signaling Technology, Massachusetts, USA), rabbit anti-octamer-binding transcription factor 4 (Oct4) (1: 1,000; #2750; Cell Signaling Technology, Massachusetts, USA), rabbit anti Oct4a (1: 500; #2890; Cell Signaling Technology, Massachusetts, USA), and rabbit anti-GAPDH (1: 2,000; #2118; Cell Signaling Technology, Massachusetts, USA) primary antibodies.

Techniques: Immunofluorescence, Western Blot