Journal: Molecular Medicine Reports

Article Title: PDCD4 promotes inflammation/fibrosis by activating the PPAR‑γ/NF‑κB pathway in mouse atrial myocytes

doi: 10.3892/mmr.2024.13333

Figure Lengend Snippet: Sequences of siPDCD4-1, siPDCD4-2 and siPDCD4-3.

Article Snippet: The HL-1 cells in the silencing group were further divided into the following four groups: i) siNC; ii) siPDCD4; iii) siPDCD4 + PPARγ inhibitor (GW9962; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. M6191-5MG; Merck KGaA); and iv) siPDCD4 + NF-κB agonist (betulinic acid; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. HY-10529; MedChemExpress).

Techniques: Sequencing

Journal: Molecular Medicine Reports

Article Title: PDCD4 promotes inflammation/fibrosis by activating the PPAR‑γ/NF‑κB pathway in mouse atrial myocytes

doi: 10.3892/mmr.2024.13333

Figure Lengend Snippet: mRNA and protein expression levels of PDCD4 in HL-1 cells after PDCD4 overexpression and silencing. (A) Transcriptional and (B) translational analysis of PDCD4 after oePDCD4 transfection by RT-qPCR and western blotting. (C) Statistical analysis of western blotting. **P<0.01, ***P<0.001 vs. oeNC. (D) Transcriptional and (E) translational analysis of PDCD4 after siPDCD4 transfection by RT-qPCR and western blotting. (F) Statistical analysis of western blotting. **P<0.01, ***P<0.001 vs. siNC. n=3. PDCD4, programmed cell death factor 4; RT-qPCR, reverse transcription-quantitative PCR; oe, overexpression; NC, negative control; si, small interfering.

Article Snippet: The HL-1 cells in the silencing group were further divided into the following four groups: i) siNC; ii) siPDCD4; iii) siPDCD4 + PPARγ inhibitor (GW9962; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. M6191-5MG; Merck KGaA); and iv) siPDCD4 + NF-κB agonist (betulinic acid; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. HY-10529; MedChemExpress).

Techniques: Expressing, Over Expression, Transfection, Quantitative RT-PCR, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Molecular Medicine Reports

Article Title: PDCD4 promotes inflammation/fibrosis by activating the PPAR‑γ/NF‑κB pathway in mouse atrial myocytes

doi: 10.3892/mmr.2024.13333

Figure Lengend Snippet: Levels of fibrosis-associated proteins and inflammatory-related cytokines in siPDCD4-HL-1 cells. (A) Western blotting indicated that the downregulated protein expression levels of collagen I, collagen III, fibronectin, α-SMA and MMP2 in si-PDCD4-HL-1 cells were attenuated by GW9662 and betulinic acid. (B) Statistical analysis of western blotting. (C) Enzyme-linked immunosorbent assays showed that the downregulated protein concentrations of IL-6, IL-17A, IFN-γ and TNF-α, and upregulated protein concentration of IL-4, in si-PDCD4-HL-1 cells were reversed by GW9662 and betulinic acid. ***P<0.001 vs. siNC; # P<0.05, ## P<0.01, ### P<0.001, vs. si-PDCD4. n=3. si, small interfering; PDCD4, programmed cell death factor 4; α-SMA, α-smooth muscle actin; MMP, matrix metalloproteinase; NC, negative control.

Article Snippet: The HL-1 cells in the silencing group were further divided into the following four groups: i) siNC; ii) siPDCD4; iii) siPDCD4 + PPARγ inhibitor (GW9962; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. M6191-5MG; Merck KGaA); and iv) siPDCD4 + NF-κB agonist (betulinic acid; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. HY-10529; MedChemExpress).

Techniques: Western Blot, Expressing, Protein Concentration, Negative Control

Journal: Molecular Medicine Reports

Article Title: PDCD4 promotes inflammation/fibrosis by activating the PPAR‑γ/NF‑κB pathway in mouse atrial myocytes

doi: 10.3892/mmr.2024.13333

Figure Lengend Snippet: Expression of PPARγ and PDCD4 in siPDCD4-HL-1. (A) Western blotting indicated that the upregulated protein expression levels of PPARγ and downregulated protein expression levels of PDCD4 in si-PDCD4-HL-1 cells were attenuated by GW9662 and betulinic acid. (B) Statistical analysis of western blotting. ***P<0.001 vs. siNC; ## P<0.01, ### P<0.001 vs. si-PDCD4. n=3. PPARγ, peroxisome proliferator-activated receptor γ; PDCD4, programmed cell death factor 4; si, small interfering; NC, negative control.

Article Snippet: The HL-1 cells in the silencing group were further divided into the following four groups: i) siNC; ii) siPDCD4; iii) siPDCD4 + PPARγ inhibitor (GW9962; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. M6191-5MG; Merck KGaA); and iv) siPDCD4 + NF-κB agonist (betulinic acid; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. HY-10529; MedChemExpress).

Techniques: Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: PDCD4 promotes inflammation/fibrosis by activating the PPAR‑γ/NF‑κB pathway in mouse atrial myocytes

doi: 10.3892/mmr.2024.13333

Figure Lengend Snippet: Expression of NF-κB subunits in siPDCD4-HL-1. (A) Western blotting showed that, compared with in the siNC group, the Nu p-p65/p65 ratio and Nu p-p50/PCNA ratio were decreased in the siPDCD4 group, and the results were partially reversed by GW9662 and betulinic acid. However, there was no significant change in Cy p-p65/p65 ratio and Cy p-p50/GAPDH ratio among the four groups. (B) Statistical analysis of western blotting. (C) Immunofluorescence analysis showed that the decreased protein expression of p-p65 in si-PDCD4-HL-1 cells was attenuated by GW9662 and betulinic acid. (D) Statistical analysis of fluorescence intensity. Magnification, ×1,000. ***P<0.001 vs. siNC; ### P<0.001 vs. si-PDCD4. n=3. si, small interfering; PDCD4, programmed cell death factor 4; NC, negative control; p-, phosphorylated; PCNA, proliferating cell nuclear antigen; Nu, nuclear; Cy, cytoplasmic.

Article Snippet: The HL-1 cells in the silencing group were further divided into the following four groups: i) siNC; ii) siPDCD4; iii) siPDCD4 + PPARγ inhibitor (GW9962; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. M6191-5MG; Merck KGaA); and iv) siPDCD4 + NF-κB agonist (betulinic acid; 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μM, cat. no. HY-10529; MedChemExpress).

Techniques: Expressing, Western Blot, Immunofluorescence, Fluorescence, Negative Control