Journal: Molecular Therapy. Nucleic Acids

Article Title: LNA-i-miR-221 activity in colorectal cancer: A reverse translational investigation

doi: 10.1016/j.omtn.2024.102221

Figure Lengend Snippet: Molecular characterization of LNA-i-miR-221-mediated effect on CRC cells (A) GSEA highlighting a modulation of the TP53 pathway on HCT 116 cells after treatment with 15 nM LNA-i-miR-221 as compared with LNA-NC-treated cells (fold change [FC] ≤ −1.5 or ≥1.5 of differentially expressed genes was considered). (B) Histogram bars showing the FC of genes responsible for TP53 pathway modulation from GEP analysis. (C) Histogram bars representing mRNA expression of miR-221 targeted genes. GAPDH was used to normalize data. (D) Protein expression of miR-221-targeted genes, GAPDH was used to normalize data; densitometric analysis of bands resulting from the western blots 48 h following treatments at the indicated conditions is reported on the right. Values ± standard deviation (SD) represented on the x axis has been normalized on GAPDH for each experimental condition and then on values from LNA-NC. (E) Curves are representative of absorbance values that indicate luciferase activity; values were normalized to SEAP to obtain the represented percentage. To calculate statistical significance, Student’s t test was applied. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: One microgram of the pEZX-MT05 vector encoding the 3′UTR region for TP53BP2, Gluc (Gaussia luciferase), and SEAP reporters (catalog No# HmiT018178-MT05), purchased from GeneCopoeia, was co-transfected with 15 nM LNA-i-miR-221 or LNA-NC and then luciferase activity was checked, according to the manufacturer’s protocol, 48 and 72 h later.

Techniques: Expressing, Western Blot, Standard Deviation, Luciferase, Activity Assay

Journal: Cell biology and toxicology

Article Title: ASPP2 deficiency attenuates lipid accumulation through the PPARγ pathway in alcoholic liver injury.

doi: 10.1007/s10565-024-09925-x

Figure Lengend Snippet: Fig. 2 ASPP2 deficiency inhibits the PPARγ and mTOR signaling path- ways. (A and B) Repre- sentative IHC analysis of PPARγ in liver sections, scar bar = 50 µm (A) and quantification analyses (B) of PPARγ positive area (%) in mouse liver tissue from wild-type mice and ASPP2- KD mice. The extended part of the black lines shows the enlarged image from the black box area. (C and D) western blot (C) and quantification analyses (D) of ASPP2, PPARγ, phospho-mTOR, phospho- S6, phospho-p70S6K, and β-actin in mouse liver tissue from wild-type mice and ASPP2-KD mice. (E and F) western blot (E) and quantification analyses (F) of ASPP2, PPARγ, phospho-mTOR, phospho- S6, phospho-p70S6K, and β-actin in primary hepatocytes (Control and ASPP2 siRNA) treated with ethanol for different times. (G and H) western blot (G) and quantification analyses (H) of ASPP2, PPARγ, phospho-mTOR, phospho-S6, phospho- p70S6K, and β-actin in primary hepatocytes (Ad- GFP and Ad-ASPP2) with the indicated treatment. The values represent the means ± SEMs (n = 6 in each group). nsP > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001. Independent- samples T tests between two groups were used for statistical analysis, and one-way ANOVA followed by Bonferroni post hoc tests for multiple comparisons were used for statistical analyses

Article Snippet: Finally, the expression of ASPP2 or PPARγ was observed by microscopy (Leica Microsystems, Mannheim, Germany) after development using a diaminobenzidine kit (Boster, Wuhan, China).

Techniques: Western Blot, Control