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Knockdown of MBD2 Attenuates LPS-Stimulated Inflammation and Apoptosis in WI-38 Cells Through the STAT-3 Pathway Cover

Knockdown of MBD2 Attenuates LPS-Stimulated Inflammation and Apoptosis in WI-38 Cells Through the STAT-3 Pathway

Archivum Immunologiae et Therapiae Experimentalis
Volume 73 (2025): Issue 1 (January 2025)
By: Yao Chen and  Liqun Lu  
Open Access
|Jul 2025

Figures & Tables

Fig 1.

MBD2 was highly expressed in LPS-stimulated WI-38 cells. (A) CCK-8 assays indicated the growth of WI-38 cells upon the treatment of LPS at the concentration of 5 μg/mL, 10 μg/mL, and 20 μg/mL for 24 h. The OD450 value was measured. (B) Immunoblot assays showed the expression of MBD2 in WI-38 cells upon the treatment of LPS for 24 h. The relative expression of MBD2 was quantified. (C) qPCR assays showed the mRNA levels of MBD2 in WI-38 cells upon the treatment of LPS for 24 h. (D) Immunoblot assays showed the expression of MBD2 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. (E) CCK-8 assays showed the growth of WI-38 cells upon the indicated treatment. The relative expression of MBD2 was quantified. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. CCK-8, cell counting kit-8; GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; LPS, lipopolysaccharide; MBD2, MBD protein 2; NC, negative control; qPCR, quantitative polymerase chain reaction.
MBD2 was highly expressed in LPS-stimulated WI-38 cells. (A) CCK-8 assays indicated the growth of WI-38 cells upon the treatment of LPS at the concentration of 5 μg/mL, 10 μg/mL, and 20 μg/mL for 24 h. The OD450 value was measured. (B) Immunoblot assays showed the expression of MBD2 in WI-38 cells upon the treatment of LPS for 24 h. The relative expression of MBD2 was quantified. (C) qPCR assays showed the mRNA levels of MBD2 in WI-38 cells upon the treatment of LPS for 24 h. (D) Immunoblot assays showed the expression of MBD2 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. (E) CCK-8 assays showed the growth of WI-38 cells upon the indicated treatment. The relative expression of MBD2 was quantified. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. CCK-8, cell counting kit-8; GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; LPS, lipopolysaccharide; MBD2, MBD protein 2; NC, negative control; qPCR, quantitative polymerase chain reaction.

Fig 2.

Knockdown of MBD2 alleviates production of cellular inflammatory cytokines in LPS-stimulated WI-38 cells. (A) qPCR assays showed the mRNA levels of TNF-α, IL-1β, and IL-6 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. (B) ELISA showed the secretion levels of TNF-α, IL-1β, and IL-6 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; MBD2, MBD protein 2; NC, negative control, qPCR, quantitative polymerase chain reaction; TNF-α, tumor necrosis factor-α.
Knockdown of MBD2 alleviates production of cellular inflammatory cytokines in LPS-stimulated WI-38 cells. (A) qPCR assays showed the mRNA levels of TNF-α, IL-1β, and IL-6 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. (B) ELISA showed the secretion levels of TNF-α, IL-1β, and IL-6 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; MBD2, MBD protein 2; NC, negative control, qPCR, quantitative polymerase chain reaction; TNF-α, tumor necrosis factor-α.

Fig 3.

Knockdown of MBD2 alleviates apoptosis in LPS-stimulated WI-38 cells. (A) Flow cytometry assays showed the apoptosis of WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. (B) Immunoblot assays showed the expression of Bax and cleaved caspase-3 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; LPS, lipopolysaccharide; MBD2, MBD protein 2; NC, negative control.
Knockdown of MBD2 alleviates apoptosis in LPS-stimulated WI-38 cells. (A) Flow cytometry assays showed the apoptosis of WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. (B) Immunoblot assays showed the expression of Bax and cleaved caspase-3 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; LPS, lipopolysaccharide; MBD2, MBD protein 2; NC, negative control.

Fig 4.

Knockdown of MBD2 regulates the STAT-3 pathway in LPS-stimulated WI-38 cells. Immunoblot assays showed the expression and phosphorylation levels of STAT-3 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; LPS, lipopolyssacharide; MBD2, MBD protein 2; NC, negative control, STAT-3, signal transducer and activator of transcription-3.
Knockdown of MBD2 regulates the STAT-3 pathway in LPS-stimulated WI-38 cells. Immunoblot assays showed the expression and phosphorylation levels of STAT-3 in WI-38 cells upon the treatment of LPS and transfection of si-NC or si-MBD2 for 24 h. *p < 0.05, **p < 0.01, ***p < 0.001, LPS vs. control, #p < 0.05, ##p < 0.01, ###p < 0.001, si-MBD2 vs. si-NC. GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; LPS, lipopolyssacharide; MBD2, MBD protein 2; NC, negative control, STAT-3, signal transducer and activator of transcription-3.
DOI: https://doi.org/10.2478/aite-2025-0021 | Journal eISSN: 1661-4917
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Language: English
Submitted on: Mar 6, 2025
|
Accepted on: May 23, 2025
|
Published on: Jul 19, 2025
Published by: Hirszfeld Institute of Immunology and Experimental Therapy
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
Pneumonia,
MBD protein 2,
Inflammatory cytokines,
Apoptosis,
STAT-3
Related subjects:
Medicine,
Basic medical science,
Biochemistry,
Immunology,
Clinical medicine,
Clinical medicine, other,
Clinical chemistry

© 2025 Yao Chen, Liqun Lu, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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