Journal: Stem cell research & therapy

Article Title: Loss of receptor tyrosine kinase-like orphan receptor 2 impairs the osteogenesis of mBMSCs by inhibiting signal transducer and activator of transcription 3.

doi: 10.1186/s13287-020-01646-2

Figure Lengend Snippet: Fig. 5 In Ror2-knockdown mBMSCs, the expression of Stat3 was decreased. a–c Quantitative real-time PCR (a) and western blot (b, c) analysis of the expression of Stat3 after osteogenic induction for 3 weeks. n = 3, *P < 0.05 vs. mBMSC-mock1. d Immunocytofluorescence staining showed the distribution of Stat3 after osteogenic induction for 24 h. Scale bar = 50 μm. e Western blot analysis revealed the expression of nuclear Stat3 and pStat3 after osteogenic induction for 24 h. f, g Overexpression of Stat3 in Ror2-knockdown mBMSCs. Stat3 protein expression was evaluated using western blot analysis. n = 3, $P < 0.05 vs. mBMSC-sh-Ror2

Article Snippet: Lentiviral vector construction and transduction The recombinant lentivirus vector downregulation of mouse Ror2 gene and overexpression of mouse Stat3 gene were generated by Cyagen Biosciences, Inc. (Guangzhou, China) and iGeneBio Biotechnology (Guangzhou, China), respectively.

Techniques: Knockdown, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Staining, Over Expression

Journal: Stem cell research & therapy

Article Title: Loss of receptor tyrosine kinase-like orphan receptor 2 impairs the osteogenesis of mBMSCs by inhibiting signal transducer and activator of transcription 3.

doi: 10.1186/s13287-020-01646-2

Figure Lengend Snippet: Fig. 6 In response to osteogenic induction, the Ror2-knockdown mBMSCs showed decreased mineralization, which was rescued by overexpression of Stat3. a The cells were stained with Alizarin Red after osteogenic induction for 3 weeks. Scale bar = 100 μm. b Alizarin Red concentrations were determined by a quantitative destaining procedure using CPC. n = 3, *P < 0.05 vs. mBMSC-mock1; $P < 0.05 vs. mBMSC- mock2; #P < 0.05 vs. mBMSC-sh-Ror2

Article Snippet: Lentiviral vector construction and transduction The recombinant lentivirus vector downregulation of mouse Ror2 gene and overexpression of mouse Stat3 gene were generated by Cyagen Biosciences, Inc. (Guangzhou, China) and iGeneBio Biotechnology (Guangzhou, China), respectively.

Techniques: Knockdown, Over Expression, Staining

Journal: Stem cell research & therapy

Article Title: Loss of receptor tyrosine kinase-like orphan receptor 2 impairs the osteogenesis of mBMSCs by inhibiting signal transducer and activator of transcription 3.

doi: 10.1186/s13287-020-01646-2

Figure Lengend Snippet: Fig. 7 In response to osteogenic induction, the less expressed osteogenic makers in Ror2-knockdown mBMSCs were restored by overexpression of Stat3. a, b Western blot analysis of the expression of osteogenic specific markers after osteogenic induction for 3 weeks. n = 3, *P < 0.05 vs. mBMSC-mock1; $P < 0.05 vs. mBMSC-mock2; #P < 0.05 vs. mBMSC-sh-Ror2

Article Snippet: Lentiviral vector construction and transduction The recombinant lentivirus vector downregulation of mouse Ror2 gene and overexpression of mouse Stat3 gene were generated by Cyagen Biosciences, Inc. (Guangzhou, China) and iGeneBio Biotechnology (Guangzhou, China), respectively.

Techniques: Knockdown, Over Expression, Western Blot, Expressing

Journal: The FEBS journal

Article Title: Long non-coding RNA RP11-820 promotes extracellular matrix production via regulating miR-3178/MYOD1 in human trabecular meshwork cells.

doi: 10.1111/febs.15058

Figure Lengend Snippet: Fig. 6. MYOD1 binds STAT3 and activates transcription of ECM genes. (A) Western blot analysis of MYOD1, Fibronectin, Collagen I and Laminin in HTMCs 1 (left) or 2 (right) transfected with the indicated condition. (B) Whole-cell lysates from MYOD1 knockdown HTMCs 1 or scramble cells were immunoprecipitated with anti-MYOD1 antibody. Precipitates were analysed by western blot analysis for MYOD1 and STAT3. (C) Whole-cell lysates from STAT3 knockdown HTMCs 1 or scramble cells were immunoprecipitated with anti-STAT3 antibody. Precipitates were analysed by western blot analysis for MYOD1 and STAT3. (D) mRNA expression of STAT3, collagen I, and fibronectin in STAT3-knockdown or scramble HTMCs 1. (E) mRNA expression of STAT3, collagen I and fibronectin in HTMCs stably expressing STAT3 or control HTMCs 1. (F, G) Western blot analysis of fibronectin, collagen I and laminin in HTMCs 1 (left) or 2 (right) transfected with the indicated condition. The values represent the mean SD of three independent experiments performed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Lentiviral expression vector generation and cell line selection To over-express RP11-820 or STAT3, the full-length RP11820 sequence or full-length STAT3 coding sequence was cloned into the PCDH-CMV-MCS-EF1-Puro lentiviral vector (Addgene, Cambridge, MA, USA) between the XbaI and EcoRI restriction sites.

Techniques: Western Blot, Transfection, Knockdown, Immunoprecipitation, Expressing, Stable Transfection, Control

Journal: The FEBS journal

Article Title: Long non-coding RNA RP11-820 promotes extracellular matrix production via regulating miR-3178/MYOD1 in human trabecular meshwork cells.

doi: 10.1111/febs.15058

Figure Lengend Snippet: Fig. 7. Proposed working model. Schematic summarizing our proposed model for lncRNA-RP11-820 regulates ECM genes in HTMCs. lncRNA-RP11-820 directly adsorb miR-3178 as a sponge, through which upregulate the expression of MYOD1. Importantly, MYOD1 can transcriptionally bind to the promoter region of ECM gene, leading to increased ECM protein deposition. Furthermore, MYOD1 can bind STAT3 to form a transcription complex, and transcriptionally activates ECM genes expression in HTMCs.

Article Snippet: Lentiviral expression vector generation and cell line selection To over-express RP11-820 or STAT3, the full-length RP11820 sequence or full-length STAT3 coding sequence was cloned into the PCDH-CMV-MCS-EF1-Puro lentiviral vector (Addgene, Cambridge, MA, USA) between the XbaI and EcoRI restriction sites.

Techniques: Expressing

Journal: BMC cancer

Article Title: Expression of STAT3 and hypoxia markers in long-term surviving malignant glioma patients.

doi: 10.1186/s12885-024-12221-w

Figure Lengend Snippet: Fig. 2 The expression of selected hypoxia-related markers (IDH1, IDH2, HIF1a, HIF1b, HIF2a, EGFR, PTEN, VEGFA, VEGFC and STAT3) in primary glioma cells GBMLe3, GBMLe4 and GBMDo2 (A) and in cryopreserved samples corresponding to the tumor used for particular primary glioma culture derivation (B) at mRNA level. The expression of mRNA was determined by RT-PCR. Data are expressed as fold increase ± SD of averages from two independent experi ments. Beta-2-microglobulin was used as a housekeeping gene. * p < 0.05 GBM26 vs. GBM43; # p < 0.05 GBM26 vs. GBM59

Article Snippet: Crispr/Cas STAT3 knockout cell model Glioma cells U87MG grown to 50–70% confluence were transfected with transfection mixture (gRNA vectors in Opti-MEM I, the donor DNA and Turbofectin 8.0 - the ratios of 3:1 for Turbofectin: DNA) as based on manufacturer’s protocol (STAT3 Human Gene Knockout Kit (CRISPR), CAT#: KN204922, Origene).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: BMC cancer

Article Title: Expression of STAT3 and hypoxia markers in long-term surviving malignant glioma patients.

doi: 10.1186/s12885-024-12221-w

Figure Lengend Snippet: Fig. 3 Comparison of tumor growth and drug accumulation in Foxn1-nu mice after implantation of glioma cell lines followed by TMZ treatment. Tumor size of implanted (n = 4) (A) U87MG IDH1wt, U87MG STAT3 KO with and without TMZ (0.9 mg/kg) treatment. Evaluation of accumulation of (B) TMZ and (C) its metabolites AIC inside the brain, tumor and plasma in tumor bearing mice with implanted U87MG IDH1wt, resp. U87MG STAT3 KO. The administration of drug (TMZ– 0.9 mg/kg) begins two weeks after implantation (from day 15. to day 28. daily). Organs were collected 15 min after last TMZ application. Confidence interval values of tumor size are shown as mean ± SD. The data of drug accumulation are expressed as ng per mg of tissue. Measurements were performed in two independent experiments

Article Snippet: Crispr/Cas STAT3 knockout cell model Glioma cells U87MG grown to 50–70% confluence were transfected with transfection mixture (gRNA vectors in Opti-MEM I, the donor DNA and Turbofectin 8.0 - the ratios of 3:1 for Turbofectin: DNA) as based on manufacturer’s protocol (STAT3 Human Gene Knockout Kit (CRISPR), CAT#: KN204922, Origene).

Techniques: Comparison, Clinical Proteomics

Journal: BMC cancer

Article Title: Expression of STAT3 and hypoxia markers in long-term surviving malignant glioma patients.

doi: 10.1186/s12885-024-12221-w

Figure Lengend Snippet: Fig. 4 The expression of selected markers related with hypoxia (IDH1, IDH2, HIF1a, HIF1b, HIF2a, EGFR, PTEN, VEGFA, VEGFC and STAT3) in glioma U87MG and U87MG STAT3 KO glioma cell lines (A) and glioma samples collected from Foxn1-nu mice with implanted U87MG and U87MG STAT3 KO glioma cells on mRNA level (B). Tumors were collected 28 days after glioma cell implantation and processed as described in Materials and methods section

Article Snippet: Crispr/Cas STAT3 knockout cell model Glioma cells U87MG grown to 50–70% confluence were transfected with transfection mixture (gRNA vectors in Opti-MEM I, the donor DNA and Turbofectin 8.0 - the ratios of 3:1 for Turbofectin: DNA) as based on manufacturer’s protocol (STAT3 Human Gene Knockout Kit (CRISPR), CAT#: KN204922, Origene).

Techniques: Expressing