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Effects of metformin on <t>cytokine</t> expression and ERK signaling <t>in</t> <t>RPMI</t> 2650 cancer cells in the presence of leptin. (A) Comparative cytokine array analysis of RPMI 2650 cancer cells treated with 10 ng/mL leptin alone and in combination with 10 mM metformin. 1=IL-2, 2=Serpin E1/PAI-1, 3=IL-18/IL-1FA. (B) Quantification of pERK/ERK expression levels in RPMI 2650 cancer cells treated with 10 ng/mL leptin and 10 ng/mL leptin+10 mM metformin. The expression of pERK and ERK was normalized to GAPDH expression. One-way ANOVA was performed (n=3). Data are presented as the group mean±standard deviation. ** P <0.01, *** P <0.005, and **** P <0.001. ERK, extracellular signal-regulated kinase; pERK, phosphorylated ERK; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IL, interleukin; PAI-1, plasminogen activator inhibitor-1.
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Effects of metformin on <t>cytokine</t> expression and ERK signaling <t>in</t> <t>RPMI</t> 2650 cancer cells in the presence of leptin. (A) Comparative cytokine array analysis of RPMI 2650 cancer cells treated with 10 ng/mL leptin alone and in combination with 10 mM metformin. 1=IL-2, 2=Serpin E1/PAI-1, 3=IL-18/IL-1FA. (B) Quantification of pERK/ERK expression levels in RPMI 2650 cancer cells treated with 10 ng/mL leptin and 10 ng/mL leptin+10 mM metformin. The expression of pERK and ERK was normalized to GAPDH expression. One-way ANOVA was performed (n=3). Data are presented as the group mean±standard deviation. ** P <0.01, *** P <0.005, and **** P <0.001. ERK, extracellular signal-regulated kinase; pERK, phosphorylated ERK; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IL, interleukin; PAI-1, plasminogen activator inhibitor-1.
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Overall study design and multiplex <t>cytokine</t> analysis of rheumatoid arthritis (RA) peripheral blood mononuclear cells (PBMCs). (A) Workflow chart outlining the overall study design, illustrating each step from patient selection to data analysis, and highlighting the methodologies used in the extraction and analysis of PBMCs from unclassified arthritis (UA), healthy controls, ACPA-negative early RA (ACPA - eRA), and ACPA-positive early RA (ACPA + eRA). (B) Dot plot showing the serum concentration levels of interferon-γ (IFN-γ) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 18), and ACPA + eRA (n = 19). The horizontal bar indicates the mean value. (C) Dot plot showing the serum concentration levels of interleukin-12 (IL-12) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 17), and ACPA + eRA (n = 19). The horizontal bar represents the mean value. (D) Scatter plot illustrating the correlation between IFN-γ and IL-12 concentrations in serum samples. Statistical significance was assessed using the Kruskal–Wallis test for (B, C) and Pearson’s correlation coefficient for (D) . P-values less than 0.05 were considered significant.
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Overall study design and multiplex <t>cytokine</t> analysis of rheumatoid arthritis (RA) peripheral blood mononuclear cells (PBMCs). (A) Workflow chart outlining the overall study design, illustrating each step from patient selection to data analysis, and highlighting the methodologies used in the extraction and analysis of PBMCs from unclassified arthritis (UA), healthy controls, ACPA-negative early RA (ACPA - eRA), and ACPA-positive early RA (ACPA + eRA). (B) Dot plot showing the serum concentration levels of interferon-γ (IFN-γ) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 18), and ACPA + eRA (n = 19). The horizontal bar indicates the mean value. (C) Dot plot showing the serum concentration levels of interleukin-12 (IL-12) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 17), and ACPA + eRA (n = 19). The horizontal bar represents the mean value. (D) Scatter plot illustrating the correlation between IFN-γ and IL-12 concentrations in serum samples. Statistical significance was assessed using the Kruskal–Wallis test for (B, C) and Pearson’s correlation coefficient for (D) . P-values less than 0.05 were considered significant.
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Effects of metformin on cytokine expression and ERK signaling in RPMI 2650 cancer cells in the presence of leptin. (A) Comparative cytokine array analysis of RPMI 2650 cancer cells treated with 10 ng/mL leptin alone and in combination with 10 mM metformin. 1=IL-2, 2=Serpin E1/PAI-1, 3=IL-18/IL-1FA. (B) Quantification of pERK/ERK expression levels in RPMI 2650 cancer cells treated with 10 ng/mL leptin and 10 ng/mL leptin+10 mM metformin. The expression of pERK and ERK was normalized to GAPDH expression. One-way ANOVA was performed (n=3). Data are presented as the group mean±standard deviation. ** P <0.01, *** P <0.005, and **** P <0.001. ERK, extracellular signal-regulated kinase; pERK, phosphorylated ERK; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IL, interleukin; PAI-1, plasminogen activator inhibitor-1.

Journal: Preventive Nutrition and Food Science

Article Title: Antitumor Effects of Metformin in Squamous Cell Carcinoma under Leptin Treatment Conditions

doi: 10.3746/pnf.2025.30.4.312

Figure Lengend Snippet: Effects of metformin on cytokine expression and ERK signaling in RPMI 2650 cancer cells in the presence of leptin. (A) Comparative cytokine array analysis of RPMI 2650 cancer cells treated with 10 ng/mL leptin alone and in combination with 10 mM metformin. 1=IL-2, 2=Serpin E1/PAI-1, 3=IL-18/IL-1FA. (B) Quantification of pERK/ERK expression levels in RPMI 2650 cancer cells treated with 10 ng/mL leptin and 10 ng/mL leptin+10 mM metformin. The expression of pERK and ERK was normalized to GAPDH expression. One-way ANOVA was performed (n=3). Data are presented as the group mean±standard deviation. ** P <0.01, *** P <0.005, and **** P <0.001. ERK, extracellular signal-regulated kinase; pERK, phosphorylated ERK; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IL, interleukin; PAI-1, plasminogen activator inhibitor-1.

Article Snippet: The supernatant from RPMI 2650 cancer cells was used in the human cytokine array kit (R&D Systems) following the manufacturer’s instructions.

Techniques: Expressing, Standard Deviation

Overall study design and multiplex cytokine analysis of rheumatoid arthritis (RA) peripheral blood mononuclear cells (PBMCs). (A) Workflow chart outlining the overall study design, illustrating each step from patient selection to data analysis, and highlighting the methodologies used in the extraction and analysis of PBMCs from unclassified arthritis (UA), healthy controls, ACPA-negative early RA (ACPA - eRA), and ACPA-positive early RA (ACPA + eRA). (B) Dot plot showing the serum concentration levels of interferon-γ (IFN-γ) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 18), and ACPA + eRA (n = 19). The horizontal bar indicates the mean value. (C) Dot plot showing the serum concentration levels of interleukin-12 (IL-12) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 17), and ACPA + eRA (n = 19). The horizontal bar represents the mean value. (D) Scatter plot illustrating the correlation between IFN-γ and IL-12 concentrations in serum samples. Statistical significance was assessed using the Kruskal–Wallis test for (B, C) and Pearson’s correlation coefficient for (D) . P-values less than 0.05 were considered significant.

Journal: Frontiers in Immunology

Article Title: Upregulation of interferon-γ response genes in monocytes and T cells identified by single-cell transcriptomics in patients with anti-citrullinated peptide antibody-positive early rheumatoid arthritis

doi: 10.3389/fimmu.2024.1439082

Figure Lengend Snippet: Overall study design and multiplex cytokine analysis of rheumatoid arthritis (RA) peripheral blood mononuclear cells (PBMCs). (A) Workflow chart outlining the overall study design, illustrating each step from patient selection to data analysis, and highlighting the methodologies used in the extraction and analysis of PBMCs from unclassified arthritis (UA), healthy controls, ACPA-negative early RA (ACPA - eRA), and ACPA-positive early RA (ACPA + eRA). (B) Dot plot showing the serum concentration levels of interferon-γ (IFN-γ) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 18), and ACPA + eRA (n = 19). The horizontal bar indicates the mean value. (C) Dot plot showing the serum concentration levels of interleukin-12 (IL-12) in UA (n = 21), controls (n = 16), ACPA - eRA (n = 17), and ACPA + eRA (n = 19). The horizontal bar represents the mean value. (D) Scatter plot illustrating the correlation between IFN-γ and IL-12 concentrations in serum samples. Statistical significance was assessed using the Kruskal–Wallis test for (B, C) and Pearson’s correlation coefficient for (D) . P-values less than 0.05 were considered significant.

Article Snippet: Concentrations of IFN-γ, IL-12 and IL-6, in serum samples of eRA patients were measured from using Millipore’s MILLIPLEX MAP High Sensitivity Human Cytokine multiplex kit (cat. no. HSTCMAG-28SK; Merck, Billerica, MA, USA) according to the manufacturer’s instructions.

Techniques: Multiplex Assay, Selection, Extraction, Concentration Assay