Journal: Applied Sciences

Article Title: Kaempferol Regresses Carcinogenesis through a Molecular Cross Talk Involved in Proliferation, Apoptosis and Inflammation on Human Cervical Cancer Cells, HeLa

doi: 10.3390/app12063155

Figure Lengend Snippet: Figure 3. Transcripts’ expression analysis (A) Graphical representation of fold change showing downregulation of various cell cycle regulatory genes, PI3K/AKT, MAPK and WNT pathways signalling molecules, while upregulation in TSGs’ expression compared to control followed by treatment with kaempferol (50 µM for 48 h). (B). Heat map demonstrating the increased and decreased expression of pro-apoptotic and anti-apoptotic genes, respectively, alongside upregulation of caspases, extrinsic receptors and ligands related to intrinsic and extrinsic pathways followed by kaempferol treatment (30 and 50 µM) at 48 h. The data are expressed as the mean ± standard deviation of three independent experiments. Statistically significant differences are marked by asterisks: Two way-ANOVA * represents p < 0.05; ** represents p < 0.01, *** represents p < 0.001.

Article Snippet: Cultured cells in complete media were exposed to 0 μM (control), 30, 40 and 50 μM of kaempferol for 48 h. DNA isolation was done in suspension buffer as per the kit’s (Quick Apoptotic DNA Ladder Detection Kit by Ray biotech) protocol, followed by resolving the DNA in 1.2% agarose gel electrophoresis.

Techniques: Expressing, Control, Standard Deviation

Journal: Applied Sciences

Article Title: Kaempferol Regresses Carcinogenesis through a Molecular Cross Talk Involved in Proliferation, Apoptosis and Inflammation on Human Cervical Cancer Cells, HeLa

doi: 10.3390/app12063155

Figure Lengend Snippet: Figure 4. (A) Graph represents modulated protein expression as fold change compared to control. Kaempferol treatment upregulated pro-apoptotic protein, whereas anti-apoptotic proteins were found to be downregulated. Images of nitrocellulose membrane showing differential protein expression of various apoptotic proteins in control and the kaempferol-treated sample (30 and 50 µM of kaempferol for 48 h). (B) Evaluation of caspase 3 and caspase 8 activity in kaempferol- treated HeLa cells at 30, 40 and 50 µM for 48 h. Graph represents increase in fold change of caspase 3, and 8 activity compared to control. (C) Fluorescent microscopic images showing TMRE fluorescence intensity reduction in a dose-dependent manner of kaempferol-treated HeLa cells (30, 40, and 50 µM for 48), signifying reduction in mitochondrial membrane potential. (D) Graphical representation of TMRE fluorescence expressed as % of kaempferol-treated HeLa cells (30, 40 and 50 µM for 48 h) in comparison to control. Data are presented as the mean ± standard deviation of three independent experiments. Two way–ANOVA * = p < 0.05; ** = p < 0.01, *** p < 0.001.

Article Snippet: Cultured cells in complete media were exposed to 0 μM (control), 30, 40 and 50 μM of kaempferol for 48 h. DNA isolation was done in suspension buffer as per the kit’s (Quick Apoptotic DNA Ladder Detection Kit by Ray biotech) protocol, followed by resolving the DNA in 1.2% agarose gel electrophoresis.

Techniques: Expressing, Control, Membrane, Activity Assay, Comparison, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enterohemorrhagic Escherichia coli O157 outer membrane vesicles administered by oral gavage cause renal tubular injury and acute kidney failure in mice

doi: 10.3389/fcimb.2025.1704731

Figure Lengend Snippet: EHEC O157 OMVs administered to mice by oral gavage cause tubular epithelial damage and apoptosis. Histopathology of paraffin sections stained with hematoxylin–eosin (A–C) and transmission electron microscopy (D–F) of the kidneys from mice treated with 100–400 µg of OMVs (A, B, D, E) and from PBS-treated mice (C, F) . (A) Tubular epithelial cell flattening (black arrows) and intraluminal detached epithelial cells (asterisks). (B) Tubular epithelial flattening (black arrows), extensive vacuolization (blue arrows), detached intraluminal epithelial cells (asterisks). (C) Normal tubules (arrows) and glomeruli in a PBS-treated mouse. (D) Microvacuolization (thin white arrows), macrovacuolization (thick white arrows), and flattening of tubular epithelial cells with a preserved brush border (white asterisks). (E) Macrovacuolization of tubular epithelial cells (red arrows) with lost brush border; the green arrow indicates an apoptotic nucleus. (F) Normal tubule in a PBS-treated mouse; the yellow arrow indicates a normal tubular epithelial cell and the white asterisk a preserved brush border. (A–C) Magnification 400x; (D–F) scale bars, 5 µm. Images are representative of findings in mice treated with 100 µg, 200 µg, or 400 µg of EHEC O157 OMVs or PBS. (G) Pathology score for tubular epithelial damage was assessed based on the extent of brush border loss, epithelial cell flattening, and vacuolization after 72 h. Data are means ± standard deviations for groups of mice administered the indicated OMV dose or PBS; **** p < 0.0001 for mice treated with 100–400 µg of OMVs compared with mice treated with PBS (one-way ANOVA). (H) Apoptosis shown as DNA laddering in the kidneys of mice treated with 100 µg (lane 2), 200 µg (lane 3), or 400 µg (lane 4) of OMVs for 72 h Lanes 1 and 5, kidneys from mice treated with 5 µg and 25 µg of OMVs, respectively. Lanes 6 and 7, kidneys from mice treated with PBS. Lane 8, kidney treated with 1 µM staurosporine (positive control). Lanes M, molecular size marker (100 bp ladder). The entire original gel is shown in .

Article Snippet: Apoptotic DNA was isolated from the pellet using the Apoptotic DNA Ladder Extraction Kit (Assay Genie, Dublin, Ireland) according to the manufacturer’s instructions.

Techniques: Histopathology, Staining, Transmission Assay, Electron Microscopy, DNA Laddering, Positive Control, Marker