Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: Primer sequences for reverse transcription-quantitative PCR.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Sequencing

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: mRNA expression analysis by RT-qPCR, and assessment of self-renewal and tumor growth in H460-derived SFCs. (A) BEP2D, H460 and A549 cells were treated with DFOG (0–20 µM) for 48 h, and cell viability was assessed using a Cell Counting Kit-8 assay. H460 and A549 cells were treated with DFOG (5 µM) for 24 h. RT-qPCR was used to evaluate the effects of DFOG (5 µM) on tumor-suppressive miRNAs, including miR-671-5p, miR-148a-3p, miR-340-5p, miR-342-3p, miR-34a-5p and miR-152-3p in (B) H460 and (C) A549 cells. (D) Comparison of miR-152-3p expression between H460 cells and H460-derived SFCs. (E) STAT3 mRNA levels and (F) p-STAT3 protein levels. Rates of (G) sphere formation and (H) colony formation (scale bar, 100 µm). Western blot analysis of (I) CD44 and CD133 expression, as well as (J) Oct4 and Sox2 expression. *P<0.05, **P<0.01 (n=3). DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR/miRNA, microRNA; p-, phosphorylated; RT-qPCR, reverse transcription-quantitative PCR; SFC, sphere-forming cell.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Expressing, Quantitative RT-PCR, Derivative Assay, Cell Counting, Comparison, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: DFOG induces miR-152-3p expression, and inhibits self-renewal and tumor growth in H460-derived SFCs. At the indicated concentrations, DFOG (A) upregulated miR-152-3p expression, and (B) decreased STAT3 mRNA expression and (C) p-STAT3 protein levels in H460-derived SFCs. (D) Sphere formation and (E) colony formation were reduced (scale bar, 100 µm). Western blot analysis showed downregulation of (F) CD44 and CD133 expression, as well as (G) Oct4 and Sox2 expression. *P<0.05, **P<0.01,***P<0.001, ****P<0.0001 (n=3). (H) Images of tumor tissue; volume quantification; weight quantification; H&E staining and immunohistochemical staining using an anti-p-STAT3 antibody (scale bar, 50 µm). Quantification of p-STAT3 protein levels and miR-152-3p levels in xenograft tumors of nude mice bearing H460-derived SFCs treated with DFOG at the indicated doses. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA; p-, phosphorylated; SFC, sphere-forming cell.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Expressing, Derivative Assay, Western Blot, Staining, Immunohistochemical staining

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: miR-152-3p mimic enhances DFOG-induced downregulation of p-STAT3 levels and inhibits self-renewal in H460-derived SFCs. Expression levels of (A) miR-152-3p and (B) STAT3 mRNA, and (C) p-STAT3 protein levels. (D) Spheres and (E) colonies formed were quantified (scale bar, 100 µm). Western blot analysis of (F) CD44 and CD133, as well as (G) Oct4 and Sox2 expression in H460-derived SFCs transfected with miR-152-3p mimic and/or treated with DFOG (5 µM). *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 (n=3). DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA; NC, negative control; p-, phosphorylated; SFC, sphere-forming cell.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Derivative Assay, Expressing, Western Blot, Transfection, Negative Control

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: miR-152-3p inhibitor antagonizes DFOG-induced suppression of p-STAT3 levels and self-renewal in H460-derived SFCs. Expression levels of (A) miR-152-3p and (B) STAT3 mRNA, and (C) p-STAT3 protein levels. (D) Spheres and (E) colonies formed were quantified (scale bar, 100 µm). Western blot analysis of (F) CD44 and CD133, as well as (G) Oct4 and Sox2 expression in H460-derived SFCs transfected with miR-152-3p inhibitor and/or treated with DFOG (5 µM). *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 (n=3). DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA; NC, negative control; p-, phosphorylated; SFC, sphere-forming cell.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Derivative Assay, Expressing, Western Blot, Transfection, Negative Control

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: STAT3 inhibitor enhances DFOG-induced suppression of self-renewal in H460-derived SFCs. Expression levels of (A) miR-152-3p and (B) STAT3 mRNA, and (C) p-STAT3 protein levels are shown. (D) Spheres and (E) colonies formed were quantified (scale bar, 100 µm). Western blot analysis of (F) CD44 and CD133, as well as (G) Oct4 and Sox2 expression in H460-derived SFCs treated with S3I 201 (10 µM) and/or DFOG (5 µM). *P<0.05, **P<0.01 and ***P<0.001 (n=3). DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA; p-, phosphorylated; SFC, sphere-forming cell.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: Effect of STAT3 overexpression and DFOG co-treatment on miR-152-3p expression. Expression levels of (A) miR-152-3p and (B) STAT3 mRNA, and (C) STAT3 protein levels in H460-derived sphere-forming cells transfected with pcDNA-STAT3 and/or treated with DFOG (5 µM). *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 (n=3). DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Over Expression, Expressing, Derivative Assay, Transfection

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: STAT3 is a direct target of miR-152-3p ( https://bibiserv.cebitec.uni-bielefeld.de/rnahybrid ). (A) Representation of the predicted miR-152-3p binding site in the 3′-UTR of STAT3 mRNA. (B) Luciferase activity of 3′-UTR-WT and 3′-UTR-MUT STAT3 3′-UTR reporters in H460 cells after transfection with the miR-152-3p mimic or miR-mimic-Cont. (C) Luciferase activity of 3′-UTR-WT STAT3 3′-UTR in H460 cells transfected with the miR-152-3p mimic and/or treated with DFOG (5 µM). *P<0.05, **P<0.01 (n=3). (D) Mechanism of action of DFOG regulating the miR-152-3p/STAT3 axis and suppressing self-renewal in non-small cell lung carcinoma. 3′-UTR, 3′-untranslated region; Cont, control; DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA; MUT, mutant; p-, phosphorylated; WT, wild-type.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Control, Mutagenesis

Journal: Oncology Reports

Article Title: 7‑Difluoromethoxyl‑5,4'‑di‑n‑octylygenistein targets the STAT3 pathway by upregulating microRNA‑152‑3p expression to inhibit self‑renewal and tumor growth in non‑small cell lung carcinoma

doi: 10.3892/or.2025.8899

Figure Lengend Snippet: DFOG induces miR-152-3p expression, and inhibits STAT3 activation and self-renewal in A549-derived SFCs. At the indicated concentrations, DFOG (A) upregulated miR-152-3p expression, and decreased (B) STAT3 mRNA expression and (C) p-STAT3 protein levels in A549-derived SFCs. (D) Spheres and (E) colonies formed were quantified (scale bar, 100 µm). Western blot analysis of (F) CD44 and CD133, as well as (G) Oct4 and Sox2 expression in A549-derived SFCs. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 (n=3). DFOG, 7-difluoromethoxyl-5,4′-di-n-octylygenistein; miR, microRNA; p-, phosphorylated; SFC, sphere-forming cell.

Article Snippet: Blocking was performed using 5% skimmed milk at 37°C for 1 h. Membranes were incubated with the primary antibody at 4°C for 6 h, and membranes were incubated with the horseradish peroxidase-conjugated IgG secondary antibody (1:1,000 dilution; cat. no. RGAR011; Proteintech Group, Inc.) at room temperature for 1 h. Antibodies against α-tubulin (1:1,000 dilution; cat. no. 2125; Cell Signaling Technology, Inc.), STAT3 (1:1,000 dilution; cat. no. 12640; Cell Signaling Technology, Inc.), phosphorylated-STAT3 (p-STAT3; 1:2,000 dilution; cat. no. 9145; Cell Signaling Technology, Inc.), CD133 (1:1,000 dilution; cat. no. 64326; Cell Signaling Technology, Inc.), CD44 (1:1,000 dilution; cat. no. 37259; Cell Signaling Technology, Inc.), Oct4 (1:1,000 dilution; cat. no. 2890; Cell Signaling Technology, Inc.) and Sox2 (1:1,000 dilution; cat. no. 3579; Cell Signaling Technology, Inc.) were used as previously described ( ).

Techniques: Expressing, Activation Assay, Derivative Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Effects of Combination Treatment with Leptin and Liraglutide on Glucose Metabolism in Insulin-Dependent Diabetic Mice

doi: 10.3390/ijms26104595

Figure Lengend Snippet: Effects of the administration of liraglutide on PTP1B expression in the arcuate nucleus (ARC) and 2DG uptake in the peripheral tissues of mice with IDDM ( A ) PTP1B mRNA expression in the ARC, ( B ) STAT3 phosphorylation in the ARC (representative samples), and ( C ) 2DG uptake in brown adipose tissue (BAT), and soleus muscle. Untreated [UNT] mice shown in panels for contrast). * p < 0.05 vs. UNT mice. ns = not significant. Significant differences between the groups are further detailed in .

Article Snippet: The sections were then incubated with anti-STAT3 phosphorylation antibody (1:100 dilution: Cell Signaling, Danvers, MA, USA) and diluted in azide blocking solution overnight at 4 °C.

Techniques: Expressing, Phospho-proteomics

Journal: Molecular Medicine Reports

Article Title: Glycoprotein 130 improves repressor element‑1 silencing transcription factor‑related axon regenerative capacity in peripheral nerves with aging

doi: 10.3892/mmr.2025.13486

Figure Lengend Snippet: Primer sequences used for RT-qPCR in the present study.

Article Snippet: Primary antibodies were as follows: rabbit polyclonal anti-REST (1:1,000, 22242-1-AP; ProteinTech), rabbit polyclonal anti-GAP43 (1:1,000, 16971-1-AP; ProteinTech), rabbit polyclonal anti-glycoprotein 130 (GP130, 1:1,000, #3732; Cell Signaling), anti-signal transducer and activator of transcription 3 (STAT3, 1:1,000, #9132; Cell Signaling), anti-phosphorylated STAT3 (pSTAT3, 1:1,000, #9131; Cell Signaling), and mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, sc-32233; Santa Cruz).

Techniques: Sequencing

Journal: Molecular Medicine Reports

Article Title: Glycoprotein 130 improves repressor element‑1 silencing transcription factor‑related axon regenerative capacity in peripheral nerves with aging

doi: 10.3892/mmr.2025.13486

Figure Lengend Snippet: Expression of molecules of the JAK1/STAT3 pathway in REST-regulated cells evaluated by RT-qPCR and western blotting. REST-OE and siREST (white bars) compared with the control (black bars). RT-qPCR analysis of: IL6 in (A) REST-OE and (B) siREST; IL6 receptor in (C) REST-OE and (D) siREST; GP130 in (E) REST-OE and (F) siREST; JAK1 in (G) REST-OE and (H) siREST; and STAT3 in (I) REST-OE and (J) siREST. Western blotting analysis of GP130 in (K) REST-OE and (L) siREST. Data are expressed as mean ± standard deviation. *P<0.05, **P<0.01 and ***P<0.001. JAK1, janus kinase 1; STAT3, signal transducer and activator of transcription 3; REST, repressor element-1 silencing transcription factor; RT-qPCR, reverse transcription-quantitative PCR; REST-OE, REST-overexpressed; siREST, REST-low expressed; GP130, glycoprotein 130.

Article Snippet: Primary antibodies were as follows: rabbit polyclonal anti-REST (1:1,000, 22242-1-AP; ProteinTech), rabbit polyclonal anti-GAP43 (1:1,000, 16971-1-AP; ProteinTech), rabbit polyclonal anti-glycoprotein 130 (GP130, 1:1,000, #3732; Cell Signaling), anti-signal transducer and activator of transcription 3 (STAT3, 1:1,000, #9132; Cell Signaling), anti-phosphorylated STAT3 (pSTAT3, 1:1,000, #9131; Cell Signaling), and mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, sc-32233; Santa Cruz).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Standard Deviation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Glycoprotein 130 improves repressor element‑1 silencing transcription factor‑related axon regenerative capacity in peripheral nerves with aging

doi: 10.3892/mmr.2025.13486

Figure Lengend Snippet: Expression of STAT3 and phosphorylation of STAT3 in REST-regulated cells evaluated by western blotting. REST-OE and siREST (white bars) compared with the control (black bars). Graphs show quantification of relative protein abundance. Western blotting analysis of STAT3 and pSTAT3 in (A) REST-OE and (B) in siREST. Ratio of phosphorylation STAT3 level for total STAT3 protein levels in (C) REST-OE and (D) in siREST. Data are presented as mean ± standard deviation. *P<0.05 and **P<0.01. STAT3, signal transducer and activator of transcription 3; REST, repressor element-1 silencing transcription factor; REST-OE, REST-overexpressed; siREST, REST-low expressed; p, phosphorylated.

Article Snippet: Primary antibodies were as follows: rabbit polyclonal anti-REST (1:1,000, 22242-1-AP; ProteinTech), rabbit polyclonal anti-GAP43 (1:1,000, 16971-1-AP; ProteinTech), rabbit polyclonal anti-glycoprotein 130 (GP130, 1:1,000, #3732; Cell Signaling), anti-signal transducer and activator of transcription 3 (STAT3, 1:1,000, #9132; Cell Signaling), anti-phosphorylated STAT3 (pSTAT3, 1:1,000, #9131; Cell Signaling), and mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, sc-32233; Santa Cruz).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Quantitative Proteomics, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Glycoprotein 130 improves repressor element‑1 silencing transcription factor‑related axon regenerative capacity in peripheral nerves with aging

doi: 10.3892/mmr.2025.13486

Figure Lengend Snippet: Summary of the expression of molecules of the JAK1/STAT3 pathway in REST-regulated cells. Despite similar changes of IL6 and IL6 receptor in REST-OE and siREST, the expression of GP130, JAK1 and pSTAT3 was increased in REST-OE and the expression of GP130, JAK1 and pSTAT3 was decreased in siREST. These findings suggest that REST regulates the expression of GAP43 by the JAK1/STAT3 pathway via the expression of GP130. (A) Summary of the expression of molecules in REST-OE. In REST-OE, the expression of IL6 was significantly increased, there was no significant difference in the expression of IL6 receptor and the expression of GP130, JAK1 and pSTAT3 were significantly decreased compared with the control. (B) Summary of the expression of molecules in siREST. In siREST, the expression of IL6 was significantly increased, there was no significant difference in the expression of IL6 receptor, and the expression of GP130, JAK1 and pSTAT3 was significantly increased compared with the control. STAT3, signal transducer and activator of transcription 3; REST, repressor element-1 silencing transcription factor; REST-OE, REST-overexpressed; siREST, REST-low expressed; p, phosphorylated; GP130, glycoprotein 130; p, phosphorylated; IL, interleukin; IL6R, IL6 receptor; GAP43, growth-associated protein 43.

Article Snippet: Primary antibodies were as follows: rabbit polyclonal anti-REST (1:1,000, 22242-1-AP; ProteinTech), rabbit polyclonal anti-GAP43 (1:1,000, 16971-1-AP; ProteinTech), rabbit polyclonal anti-glycoprotein 130 (GP130, 1:1,000, #3732; Cell Signaling), anti-signal transducer and activator of transcription 3 (STAT3, 1:1,000, #9132; Cell Signaling), anti-phosphorylated STAT3 (pSTAT3, 1:1,000, #9131; Cell Signaling), and mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, sc-32233; Santa Cruz).

Techniques: Expressing, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Solid stress compression enhances breast cancer cell migration through the upregulation of Interleukin-6

doi: 10.3389/fcell.2025.1541953

Figure Lengend Snippet: Incremental solid stress compression upregulates protein levels of IL-6 and SNAI1 as well as secretion of IL-6 in a biphasic manner (A) Representative western blots of cell lysates harvested from MDA-MB-231 cells under 16 h of incremental solid stress compression using IL-6 and SNAI1 antibodies. GAPDH was used as loading control (B) Densitometry quantification of IL-6 protein levels using Image Lab. Bar graph represents the mean relative fold changes compared to uncompressed across three independent experiments (n = 3). Error bars represent standard deviation. Statistical analysis was performed using 2-tailed student T-test (*) p ≤ 0.05, (**) p ≤ 0.01 (C) Densitometry quantification of SNAI1 protein level using Image Lab. Bar graph represents the mean relative fold changes compared to uncompressed across three independent experiments (n = 3). Error bars represent standard deviation. Statistical analysis was performed using 2-tailed student T-test, (*) p ≤ 0.05, (**) p ≤ 0.01. (D) (Top) Representative Western blot of conditioned media harvested after 16 h of incremental solid stress compression and probed with anti-IL-6 antibody. (Bottom) Representative Ponceau S Staining highlighting equal protein loading of the conditioned media. (E) Densitometry quantification of IL-6 cytokine secretion using Image Lab. Bar graph represents the mean relative fold changes compared to uncompressed across three independent experiments (n = 3). Error bars represent standard deviation. Statistical analysis was performed using 2-tailed student T-test, (*) p ≤ 0.05 and (N.S) Not Significant. (F) Representative Western blot image using anti-phospho-STAT3 (pSTAT3) at Tyr-705 antibody for MDA-MB-231 cells under 16 h of incremental solid stress compression. (G) Densitometry quantification of pSTAT3 using Image Lab. Bar graph represents the mean relative fold changes compared to uncompressed across three independent experiments (n = 3). Error bars represent standard deviation. Statistical analysis was performed using 2-tailed student T-test, (*) p ≤ 0.05 and (N.S) Not Significant.

Article Snippet: Phosphorylated STAT3 Tyrosine-705 (D3A7) XP 9145 , Cell Signaling , Rabbit , 1:1000 in 5% BSA.

Techniques: Western Blot, Control, Standard Deviation, Staining