Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly reduces melanoma cell viability and proliferation. STAT1 mRNA expression was analyzed in (A) A375 and (B) RPMI-7951 melanoma cells post-STAT1 knockdown using siRNA. (C) Western blotting was used to assess STAT1 protein levels in melanoma cell lines following siRNA-mediated knockdown. Cell viability was evaluated after STAT1 knockdown in (D) A375 and (E) RPMI-7951 cells using Cell Counting Kit-8 assays. (F) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown. (G) Quantification of colony formation assay results. **P<0.01 vs. si-NC group. siRNA, small interfering RNA; STAT1, signal transducer and activator of transcription 1; si-STAT1, siRNA targeting STAT1; si-NC, negative control siRNA.

Article Snippet: Melanoma cells (A375 and RPMI-7951) were seeded into 6-well plates (cat. no. 3516; Corning, Inc.) at a density of 500–1,000 cells per well and allowed to adhere for 24 h. Cells were then transfected with si-NC (cat. no. A09010) or si-STAT1 (cat. no. A09009; GenePharma), as well as transfected using a lentiviral vector for TUBB4A overexpression (Ov-TUBB4A; pReceiver-Lv105; GeneCopoeia, Inc.) or Ov-NC (pReceiver-Lv105 Empty Vector; GeneCopoeia, Inc.).

Techniques: Knockdown, Expressing, Western Blot, Cell Counting, Colony Assay, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly promotes melanoma cell apoptosis and inhibits migration. (A and B) Apoptosis was assessed and quantified in A375 and RPMI-7951 cells following STAT1 knockdown using flow cytometry. (C) Transwell migration assays were conducted to evaluate cell migration following STAT1 knockdown. Magnification, ×200. (D) Quantification of migration capacity in A375 and RPMI-7951 cells following STAT1 knockdown. *P<0.05 vs. si-NC. STAT1, signal transducer and activator of transcription 1; si-STAT1, small interfering RNA targeting STAT1; si-NC, negative control small interfering RNA.

Article Snippet: Melanoma cells (A375 and RPMI-7951) were seeded into 6-well plates (cat. no. 3516; Corning, Inc.) at a density of 500–1,000 cells per well and allowed to adhere for 24 h. Cells were then transfected with si-NC (cat. no. A09010) or si-STAT1 (cat. no. A09009; GenePharma), as well as transfected using a lentiviral vector for TUBB4A overexpression (Ov-TUBB4A; pReceiver-Lv105; GeneCopoeia, Inc.) or Ov-NC (pReceiver-Lv105 Empty Vector; GeneCopoeia, Inc.).

Techniques: Knockdown, Migration, Flow Cytometry, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 regulates TUBB4A expression at the transcription level. (A) STAT1 mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (B) TUBB4A mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (C) Specific fragments of the TUBB4A promoter region were cloned into the luciferase reporter plasmids upstream of the firefly luciferase gene. (D) Transcriptional activity of various TUBB4A promoter fragments was analyzed by luciferase reporter assay in 293T cells, with the −1,783 and −1,771 fragments exhibiting the highest activity. (E) STAT1 siRNA-mediated knockdown significantly reduced STAT1 mRNA levels in A375 cells. (F) STAT1 knockdown significantly reduced the luciferase activity of the −1,783 fragment of the TUBB4A promoter, but not the −1,771 fragment. (G) Chromatin immunoprecipitation assays were performed in A375 and RPMI-7951 cells targeting the −1,783 binding site in the TUBB4A promoter region. Quantitative PCR provided evidence of STAT1 binding to this region. Genomic DNA input was set to 100%. **P<0.01 vs. si-NC; ## P<0.01 vs. PGL3; && P<0.01 vs. IgG. STAT1, signal transducer and activator of transcription 1; siRNA, small interfering RNA; si-NC, negative control siRNA; si-STAT1, siRNA targeting STAT1; si-STAT1-1, siRNA targeting STAT1 sequence 1; si-STAT1-2, siRNA targeting STAT1 sequence 2; TUBB4A, tubulin β4A; PGL3, promoter-gluc luciferase 3; LUC, firefly luciferase gene.

Article Snippet: Melanoma cells (A375 and RPMI-7951) were seeded into 6-well plates (cat. no. 3516; Corning, Inc.) at a density of 500–1,000 cells per well and allowed to adhere for 24 h. Cells were then transfected with si-NC (cat. no. A09010) or si-STAT1 (cat. no. A09009; GenePharma), as well as transfected using a lentiviral vector for TUBB4A overexpression (Ov-TUBB4A; pReceiver-Lv105; GeneCopoeia, Inc.) or Ov-NC (pReceiver-Lv105 Empty Vector; GeneCopoeia, Inc.).

Techniques: Expressing, Knockdown, Transfection, Clone Assay, Luciferase, Activity Assay, Reporter Assay, Chromatin Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction, Small Interfering RNA, Negative Control, Sequencing

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression mitigates the effects of STAT1 knockdown on cell viability and proliferation. TUBB4A mRNA levels were measured in (A) A375 and (B) RPMI-7951 cells following TUBB4A overexpression mediated by a lentiviral vector. (C) TUBB4A protein expression was analyzed after its overexpression. Combined STAT1 knockdown and TUBB4A overexpression transfections were performed, followed by a western blot analysis of TUBB4A protein levels in (D) A375 and (E) RPMI-7951 cells. Cell viability was assessed via Cell Counting Kit-8 assays following combined STAT1 knockdown and TUBB4A overexpression in (F) A375 and (G) RPMI-7951 cells. (H) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown and TUBB4A overexpression. (I) Quantification of colony formation assay results. **P<0.01 vs. Ov-NC; ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; STAT1, signal transducer and activator of transcription 1; si-NC, negative control small interfering RNA; si-STAT1, small interfering RNA targeting STAT1; Ov-NC, negative control lentiviral overexpression vector; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: Melanoma cells (A375 and RPMI-7951) were seeded into 6-well plates (cat. no. 3516; Corning, Inc.) at a density of 500–1,000 cells per well and allowed to adhere for 24 h. Cells were then transfected with si-NC (cat. no. A09010) or si-STAT1 (cat. no. A09009; GenePharma), as well as transfected using a lentiviral vector for TUBB4A overexpression (Ov-TUBB4A; pReceiver-Lv105; GeneCopoeia, Inc.) or Ov-NC (pReceiver-Lv105 Empty Vector; GeneCopoeia, Inc.).

Techniques: Over Expression, Knockdown, Plasmid Preparation, Expressing, Transfection, Western Blot, Cell Counting, Colony Assay, Negative Control, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression reverses the effects of STAT1 knockdown on apoptosis, migration and tumor growth. (A-D) Apoptosis and migration were evaluated in A375 and RPMI-7951 cells following STAT1 knockdown and TUBB4A overexpression. Magnification, ×200. (E) Representative images of isolated xenograft tumors in mice. Tumor volumes were measured in nude mice injected subcutaneously with 2×10 6 A375 cells that had been subject to STAT1 knockdown and TUBB4A overexpression. (F) Quantification of mouse tumor volumes showed that STAT1 knockdown significantly suppressed tumor growth, whereas TUBB4A overexpression reversed this inhibitory effect. ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; si-NC, negative control small interfering RNA; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: Melanoma cells (A375 and RPMI-7951) were seeded into 6-well plates (cat. no. 3516; Corning, Inc.) at a density of 500–1,000 cells per well and allowed to adhere for 24 h. Cells were then transfected with si-NC (cat. no. A09010) or si-STAT1 (cat. no. A09009; GenePharma), as well as transfected using a lentiviral vector for TUBB4A overexpression (Ov-TUBB4A; pReceiver-Lv105; GeneCopoeia, Inc.) or Ov-NC (pReceiver-Lv105 Empty Vector; GeneCopoeia, Inc.).

Techniques: Over Expression, Knockdown, Migration, Isolation, Injection, Negative Control, Small Interfering RNA, Plasmid Preparation

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were blocked with 5 % non-fat milk in TBS-T and incubated overnight at 4 °C with primary antibodies against phospho-AKT (Thr308), phospho-NFκB p65 (Ser536), phospho-p38 MAPK (Thr180/Tyr182), phospho-STAT1 (Tyr701), and phospho-STAT3 (Tyr705) (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 and TUBB4A expression levels are significantly correlated and upregulated in melanoma. (A) Analysis of data in the Gene Expression Profiling Interactive Analysis 2 database revealed a significant elevation of STAT1 expression in melanoma tissues. Quantification of (B) STAT1 and (C) TUBB4A mRNA levels in 31 paired melanoma and normal tissue samples from patients with SKCM demonstrated a significant upregulation of both STAT1 and TUBB4A expression in melanoma tissues. (D) Correlation analysis of STAT1 and TUBB4A mRNA levels in melanoma samples showed a significant positive correlation. Comparative analysis of (E) STAT1 and (F) TUBB4A mRNA expression in normal HEM and melanoma cell lines revealed significantly higher expression in the melanoma cell lines. *P<0.05 between groups; **P<0.01 compared with HEM cells. HEM, human epidermal melanocytes; STAT1, signal transducer and activator of transcription 1; TUBB4A, tubulin β4A; TPM, transcripts per million; SKCM, skin cutaneous melanoma.

Article Snippet: For each reaction, 100 μg chromatin lysate was diluted to a final volume of 500 μl with ChIP dilution buffer and incubated with 5 μg of anti-STAT1 antibodies (1:100; cat. no. 14994; Cell Signaling Technology, Inc.) or control rabbit IgG (1:100; cat. no. 2729; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Expressing, Gene Expression

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly reduces melanoma cell viability and proliferation. STAT1 mRNA expression was analyzed in (A) A375 and (B) RPMI-7951 melanoma cells post-STAT1 knockdown using siRNA. (C) Western blotting was used to assess STAT1 protein levels in melanoma cell lines following siRNA-mediated knockdown. Cell viability was evaluated after STAT1 knockdown in (D) A375 and (E) RPMI-7951 cells using Cell Counting Kit-8 assays. (F) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown. (G) Quantification of colony formation assay results. **P<0.01 vs. si-NC group. siRNA, small interfering RNA; STAT1, signal transducer and activator of transcription 1; si-STAT1, siRNA targeting STAT1; si-NC, negative control siRNA.

Article Snippet: For each reaction, 100 μg chromatin lysate was diluted to a final volume of 500 μl with ChIP dilution buffer and incubated with 5 μg of anti-STAT1 antibodies (1:100; cat. no. 14994; Cell Signaling Technology, Inc.) or control rabbit IgG (1:100; cat. no. 2729; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Knockdown, Expressing, Western Blot, Cell Counting, Colony Assay, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly promotes melanoma cell apoptosis and inhibits migration. (A and B) Apoptosis was assessed and quantified in A375 and RPMI-7951 cells following STAT1 knockdown using flow cytometry. (C) Transwell migration assays were conducted to evaluate cell migration following STAT1 knockdown. Magnification, ×200. (D) Quantification of migration capacity in A375 and RPMI-7951 cells following STAT1 knockdown. *P<0.05 vs. si-NC. STAT1, signal transducer and activator of transcription 1; si-STAT1, small interfering RNA targeting STAT1; si-NC, negative control small interfering RNA.

Article Snippet: For each reaction, 100 μg chromatin lysate was diluted to a final volume of 500 μl with ChIP dilution buffer and incubated with 5 μg of anti-STAT1 antibodies (1:100; cat. no. 14994; Cell Signaling Technology, Inc.) or control rabbit IgG (1:100; cat. no. 2729; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Knockdown, Migration, Flow Cytometry, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 regulates TUBB4A expression at the transcription level. (A) STAT1 mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (B) TUBB4A mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (C) Specific fragments of the TUBB4A promoter region were cloned into the luciferase reporter plasmids upstream of the firefly luciferase gene. (D) Transcriptional activity of various TUBB4A promoter fragments was analyzed by luciferase reporter assay in 293T cells, with the −1,783 and −1,771 fragments exhibiting the highest activity. (E) STAT1 siRNA-mediated knockdown significantly reduced STAT1 mRNA levels in A375 cells. (F) STAT1 knockdown significantly reduced the luciferase activity of the −1,783 fragment of the TUBB4A promoter, but not the −1,771 fragment. (G) Chromatin immunoprecipitation assays were performed in A375 and RPMI-7951 cells targeting the −1,783 binding site in the TUBB4A promoter region. Quantitative PCR provided evidence of STAT1 binding to this region. Genomic DNA input was set to 100%. **P<0.01 vs. si-NC; ## P<0.01 vs. PGL3; && P<0.01 vs. IgG. STAT1, signal transducer and activator of transcription 1; siRNA, small interfering RNA; si-NC, negative control siRNA; si-STAT1, siRNA targeting STAT1; si-STAT1-1, siRNA targeting STAT1 sequence 1; si-STAT1-2, siRNA targeting STAT1 sequence 2; TUBB4A, tubulin β4A; PGL3, promoter-gluc luciferase 3; LUC, firefly luciferase gene.

Article Snippet: For each reaction, 100 μg chromatin lysate was diluted to a final volume of 500 μl with ChIP dilution buffer and incubated with 5 μg of anti-STAT1 antibodies (1:100; cat. no. 14994; Cell Signaling Technology, Inc.) or control rabbit IgG (1:100; cat. no. 2729; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Expressing, Knockdown, Transfection, Clone Assay, Luciferase, Activity Assay, Reporter Assay, Chromatin Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction, Small Interfering RNA, Negative Control, Sequencing

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression mitigates the effects of STAT1 knockdown on cell viability and proliferation. TUBB4A mRNA levels were measured in (A) A375 and (B) RPMI-7951 cells following TUBB4A overexpression mediated by a lentiviral vector. (C) TUBB4A protein expression was analyzed after its overexpression. Combined STAT1 knockdown and TUBB4A overexpression transfections were performed, followed by a western blot analysis of TUBB4A protein levels in (D) A375 and (E) RPMI-7951 cells. Cell viability was assessed via Cell Counting Kit-8 assays following combined STAT1 knockdown and TUBB4A overexpression in (F) A375 and (G) RPMI-7951 cells. (H) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown and TUBB4A overexpression. (I) Quantification of colony formation assay results. **P<0.01 vs. Ov-NC; ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; STAT1, signal transducer and activator of transcription 1; si-NC, negative control small interfering RNA; si-STAT1, small interfering RNA targeting STAT1; Ov-NC, negative control lentiviral overexpression vector; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: For each reaction, 100 μg chromatin lysate was diluted to a final volume of 500 μl with ChIP dilution buffer and incubated with 5 μg of anti-STAT1 antibodies (1:100; cat. no. 14994; Cell Signaling Technology, Inc.) or control rabbit IgG (1:100; cat. no. 2729; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Over Expression, Knockdown, Plasmid Preparation, Expressing, Transfection, Western Blot, Cell Counting, Colony Assay, Negative Control, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression reverses the effects of STAT1 knockdown on apoptosis, migration and tumor growth. (A-D) Apoptosis and migration were evaluated in A375 and RPMI-7951 cells following STAT1 knockdown and TUBB4A overexpression. Magnification, ×200. (E) Representative images of isolated xenograft tumors in mice. Tumor volumes were measured in nude mice injected subcutaneously with 2×10 6 A375 cells that had been subject to STAT1 knockdown and TUBB4A overexpression. (F) Quantification of mouse tumor volumes showed that STAT1 knockdown significantly suppressed tumor growth, whereas TUBB4A overexpression reversed this inhibitory effect. ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; si-NC, negative control small interfering RNA; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: For each reaction, 100 μg chromatin lysate was diluted to a final volume of 500 μl with ChIP dilution buffer and incubated with 5 μg of anti-STAT1 antibodies (1:100; cat. no. 14994; Cell Signaling Technology, Inc.) or control rabbit IgG (1:100; cat. no. 2729; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Over Expression, Knockdown, Migration, Isolation, Injection, Negative Control, Small Interfering RNA, Plasmid Preparation

Journal: Journal of Advanced Research

Article Title: Transglutaminase 2 modulates inflammatory angiogenesis via vascular endothelial growth factor receptor 2 pathway in inflammatory bowel disease

doi: 10.1016/j.jare.2025.07.002

Figure Lengend Snippet: STAT1-TGM2-VEGFR2 axis drives inflammatory angiogenesis in IBD. (A) The mRNA transcription of TGM2 was significantly upregulated upon stimulation with inflammatory cytokines IL-9, IL-23, and IFN-γ in HIMECs (n = 3 in each group). (B, C) The results of WB demonstrated that stimulation with IL-9, IL-23, and IFN-γ upregulated the protein expression of TGM2 in HIMEC. (D) Stimulation with inflammatory cytokines IL-9, IL-23, and IFN-γ significantly augmented the in vitro angiogenic capacity of HIMECs, while knockdown of TGM2 effectively reversed this effect. (E, F) IL-9, IL-23 and IFN-γ stimulation significantly enhanced the phosphorylation of VEGFR2 at Tyr1059, Tyr1214 and activated the downstream pathways such as FAK, PLC-γ and MAPK pathways in HIMEC, which were reversed by TGM2 knockdown. (G) IF results showed VEGFR2 co-localizes with TGM2 in HIMEC, mainly on the cell membrane. (H) Co-IP results substantiated the interaction between TGM2 and VEGFR2. (I) The CHIP assay confirmed the binding of STAT1 to the upstream promoter region of TGM2, thereby facilitating transcription following IFN-γ stimulation. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The antibody-protein complexes were immunoprecipitated using an anti-p-STAT1 antibody (9107, CST, USA) and control IgG.

Techniques: Expressing, In Vitro, Knockdown, Phospho-proteomics, Membrane, Co-Immunoprecipitation Assay, Binding Assay

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 and TUBB4A expression levels are significantly correlated and upregulated in melanoma. (A) Analysis of data in the Gene Expression Profiling Interactive Analysis 2 database revealed a significant elevation of STAT1 expression in melanoma tissues. Quantification of (B) STAT1 and (C) TUBB4A mRNA levels in 31 paired melanoma and normal tissue samples from patients with SKCM demonstrated a significant upregulation of both STAT1 and TUBB4A expression in melanoma tissues. (D) Correlation analysis of STAT1 and TUBB4A mRNA levels in melanoma samples showed a significant positive correlation. Comparative analysis of (E) STAT1 and (F) TUBB4A mRNA expression in normal HEM and melanoma cell lines revealed significantly higher expression in the melanoma cell lines. *P<0.05 between groups; **P<0.01 compared with HEM cells. HEM, human epidermal melanocytes; STAT1, signal transducer and activator of transcription 1; TUBB4A, tubulin β4A; TPM, transcripts per million; SKCM, skin cutaneous melanoma.

Article Snippet: Membranes were blocked with 5% non-fat milk (cat. no. 1706404; Bio-Rad Laboratories, Inc.) in TBST with 0.1% Tween 20 for 1 h at 25°C and incubated overnight at 4°C with primary antibodies against STAT1 (1:1,000; cat. no. 14994; Cell Signaling Technology, Inc.) and TUBB4A (1:1,000; cat. no. ab11315; Abcam), as well as GAPDH (1:5,000; cat. no. 2118; Cell Signaling Technology, Inc.) serving as a loading control.

Techniques: Expressing, Gene Expression

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly reduces melanoma cell viability and proliferation. STAT1 mRNA expression was analyzed in (A) A375 and (B) RPMI-7951 melanoma cells post-STAT1 knockdown using siRNA. (C) Western blotting was used to assess STAT1 protein levels in melanoma cell lines following siRNA-mediated knockdown. Cell viability was evaluated after STAT1 knockdown in (D) A375 and (E) RPMI-7951 cells using Cell Counting Kit-8 assays. (F) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown. (G) Quantification of colony formation assay results. **P<0.01 vs. si-NC group. siRNA, small interfering RNA; STAT1, signal transducer and activator of transcription 1; si-STAT1, siRNA targeting STAT1; si-NC, negative control siRNA.

Article Snippet: Membranes were blocked with 5% non-fat milk (cat. no. 1706404; Bio-Rad Laboratories, Inc.) in TBST with 0.1% Tween 20 for 1 h at 25°C and incubated overnight at 4°C with primary antibodies against STAT1 (1:1,000; cat. no. 14994; Cell Signaling Technology, Inc.) and TUBB4A (1:1,000; cat. no. ab11315; Abcam), as well as GAPDH (1:5,000; cat. no. 2118; Cell Signaling Technology, Inc.) serving as a loading control.

Techniques: Knockdown, Expressing, Western Blot, Cell Counting, Colony Assay, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly promotes melanoma cell apoptosis and inhibits migration. (A and B) Apoptosis was assessed and quantified in A375 and RPMI-7951 cells following STAT1 knockdown using flow cytometry. (C) Transwell migration assays were conducted to evaluate cell migration following STAT1 knockdown. Magnification, ×200. (D) Quantification of migration capacity in A375 and RPMI-7951 cells following STAT1 knockdown. *P<0.05 vs. si-NC. STAT1, signal transducer and activator of transcription 1; si-STAT1, small interfering RNA targeting STAT1; si-NC, negative control small interfering RNA.

Article Snippet: Membranes were blocked with 5% non-fat milk (cat. no. 1706404; Bio-Rad Laboratories, Inc.) in TBST with 0.1% Tween 20 for 1 h at 25°C and incubated overnight at 4°C with primary antibodies against STAT1 (1:1,000; cat. no. 14994; Cell Signaling Technology, Inc.) and TUBB4A (1:1,000; cat. no. ab11315; Abcam), as well as GAPDH (1:5,000; cat. no. 2118; Cell Signaling Technology, Inc.) serving as a loading control.

Techniques: Knockdown, Migration, Flow Cytometry, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 regulates TUBB4A expression at the transcription level. (A) STAT1 mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (B) TUBB4A mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (C) Specific fragments of the TUBB4A promoter region were cloned into the luciferase reporter plasmids upstream of the firefly luciferase gene. (D) Transcriptional activity of various TUBB4A promoter fragments was analyzed by luciferase reporter assay in 293T cells, with the −1,783 and −1,771 fragments exhibiting the highest activity. (E) STAT1 siRNA-mediated knockdown significantly reduced STAT1 mRNA levels in A375 cells. (F) STAT1 knockdown significantly reduced the luciferase activity of the −1,783 fragment of the TUBB4A promoter, but not the −1,771 fragment. (G) Chromatin immunoprecipitation assays were performed in A375 and RPMI-7951 cells targeting the −1,783 binding site in the TUBB4A promoter region. Quantitative PCR provided evidence of STAT1 binding to this region. Genomic DNA input was set to 100%. **P<0.01 vs. si-NC; ## P<0.01 vs. PGL3; && P<0.01 vs. IgG. STAT1, signal transducer and activator of transcription 1; siRNA, small interfering RNA; si-NC, negative control siRNA; si-STAT1, siRNA targeting STAT1; si-STAT1-1, siRNA targeting STAT1 sequence 1; si-STAT1-2, siRNA targeting STAT1 sequence 2; TUBB4A, tubulin β4A; PGL3, promoter-gluc luciferase 3; LUC, firefly luciferase gene.

Article Snippet: Membranes were blocked with 5% non-fat milk (cat. no. 1706404; Bio-Rad Laboratories, Inc.) in TBST with 0.1% Tween 20 for 1 h at 25°C and incubated overnight at 4°C with primary antibodies against STAT1 (1:1,000; cat. no. 14994; Cell Signaling Technology, Inc.) and TUBB4A (1:1,000; cat. no. ab11315; Abcam), as well as GAPDH (1:5,000; cat. no. 2118; Cell Signaling Technology, Inc.) serving as a loading control.

Techniques: Expressing, Knockdown, Transfection, Clone Assay, Luciferase, Activity Assay, Reporter Assay, Chromatin Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction, Small Interfering RNA, Negative Control, Sequencing

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression mitigates the effects of STAT1 knockdown on cell viability and proliferation. TUBB4A mRNA levels were measured in (A) A375 and (B) RPMI-7951 cells following TUBB4A overexpression mediated by a lentiviral vector. (C) TUBB4A protein expression was analyzed after its overexpression. Combined STAT1 knockdown and TUBB4A overexpression transfections were performed, followed by a western blot analysis of TUBB4A protein levels in (D) A375 and (E) RPMI-7951 cells. Cell viability was assessed via Cell Counting Kit-8 assays following combined STAT1 knockdown and TUBB4A overexpression in (F) A375 and (G) RPMI-7951 cells. (H) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown and TUBB4A overexpression. (I) Quantification of colony formation assay results. **P<0.01 vs. Ov-NC; ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; STAT1, signal transducer and activator of transcription 1; si-NC, negative control small interfering RNA; si-STAT1, small interfering RNA targeting STAT1; Ov-NC, negative control lentiviral overexpression vector; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: Membranes were blocked with 5% non-fat milk (cat. no. 1706404; Bio-Rad Laboratories, Inc.) in TBST with 0.1% Tween 20 for 1 h at 25°C and incubated overnight at 4°C with primary antibodies against STAT1 (1:1,000; cat. no. 14994; Cell Signaling Technology, Inc.) and TUBB4A (1:1,000; cat. no. ab11315; Abcam), as well as GAPDH (1:5,000; cat. no. 2118; Cell Signaling Technology, Inc.) serving as a loading control.

Techniques: Over Expression, Knockdown, Plasmid Preparation, Expressing, Transfection, Western Blot, Cell Counting, Colony Assay, Negative Control, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression reverses the effects of STAT1 knockdown on apoptosis, migration and tumor growth. (A-D) Apoptosis and migration were evaluated in A375 and RPMI-7951 cells following STAT1 knockdown and TUBB4A overexpression. Magnification, ×200. (E) Representative images of isolated xenograft tumors in mice. Tumor volumes were measured in nude mice injected subcutaneously with 2×10 6 A375 cells that had been subject to STAT1 knockdown and TUBB4A overexpression. (F) Quantification of mouse tumor volumes showed that STAT1 knockdown significantly suppressed tumor growth, whereas TUBB4A overexpression reversed this inhibitory effect. ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; si-NC, negative control small interfering RNA; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: Membranes were blocked with 5% non-fat milk (cat. no. 1706404; Bio-Rad Laboratories, Inc.) in TBST with 0.1% Tween 20 for 1 h at 25°C and incubated overnight at 4°C with primary antibodies against STAT1 (1:1,000; cat. no. 14994; Cell Signaling Technology, Inc.) and TUBB4A (1:1,000; cat. no. ab11315; Abcam), as well as GAPDH (1:5,000; cat. no. 2118; Cell Signaling Technology, Inc.) serving as a loading control.

Techniques: Over Expression, Knockdown, Migration, Isolation, Injection, Negative Control, Small Interfering RNA, Plasmid Preparation

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Therapeutic effect of eravacycline against carbapenem-resistant hypervirulent Klebsiella pneumoniae in mouse models

doi: 10.1128/aac.01237-25

Figure Lengend Snippet: Expression and activation of STAT1 in mouse lung tissues. ( A ) mRNA expression levels of STAT1 in the lung tissues of mice across the uninfected control, the infection control, and the treatment groups. ( B ) Protein expression levels of p-STAT1 in the uninfected control, the infection control, and the treatment groups of mice. Data are shown as mean ± standard error of the mean, n = 3. Hashtags (#) represent the infection control group compared to the uninfected control group, and asterisks (*) represent the infection control group compared to the corresponding treatment group (* P < 0.05, ** P < 0.01, and ### P < 0.001).

Article Snippet: The membrane was blocked with TBST buffer (G2150, Servicebio Technology Co., Ltd. Wuhan, China) containing 5% bovine serum albumin and then incubated with primary antibody STAT1 (1:1,000, #9172T, Cell Signaling Technology, MA, USA), p-STAT1 (1:1,000, #7649T, Cell Signaling Technology, MA, USA), and β-actin (1:10,000, AC004, Abclonal, Wuhan, China) overnight at 4°C.

Techniques: Expressing, Activation Assay, Control, Infection

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Therapeutic effect of eravacycline against carbapenem-resistant hypervirulent Klebsiella pneumoniae in mouse models

doi: 10.1128/aac.01237-25

Figure Lengend Snippet: Expression and activation of STAT1 in mouse lung tissues. ( A ) mRNA expression levels of STAT1 in the lung tissues of mice across the uninfected control, the infection control, and the treatment groups. ( B ) Protein expression levels of p-STAT1 in the uninfected control, the infection control, and the treatment groups of mice. Data are shown as mean ± standard error of the mean, n = 3. Hashtags (#) represent the infection control group compared to the uninfected control group, and asterisks (*) represent the infection control group compared to the corresponding treatment group (* P < 0.05, ** P < 0.01, and ### P < 0.001).

Article Snippet: The membrane was blocked with TBST buffer (G2150, Servicebio Technology Co., Ltd. Wuhan, China) containing 5% bovine serum albumin and then incubated with primary antibody STAT1 (1:1,000, #9172T, Cell Signaling Technology, MA, USA), p-STAT1 (1:1,000, #7649T, Cell Signaling Technology, MA, USA), and β-actin (1:10,000, AC004, Abclonal, Wuhan, China) overnight at 4°C.

Techniques: Expressing, Activation Assay, Control, Infection