Images and Tabular Data Supporting: "Aceh Patchouli Oil (Pogostemon cablin) Inducing Apoptosis of HeLa Cervical Cancer Cells Through Modulation of the p-AKT Pathway"
Description
This dataset contains all primary figures and tabular data associated with the research article investigating the anticancer activity of Aceh patchouli oil (Pogostemon cablin) against HeLa cervical cancer cells, with emphasis on apoptosis induction via modulation of the phosphorylated AKT (p-AKT) signaling pathway. The dataset includes the following: - Figures (Images): Cell Morphology Images (Before and After Treatment); this section contains brightfield microscopy images documenting morphological changes in HeLa cervical cancer cells before (untreated control) and after treatment with Aceh patchouli oil at various concentrations, Cytotoxicity assay results (CCK-8 assay); The bar graph illustrates a dose-dependent reduction in cell viability (expressed as percentage relative to untreated control, %), with the x-axis representing treatment concentrations and the y-axis representing percentage cell viability, Flow cytometry; dot plots (Annexin V/PI staining) depicting apoptotic and necrotic cell populations following treatment with patchouli oil at various concentrations and This section contains flow cytometry data and representative dot plot graphs from Annexin V/Propidium Iodide (PI) dual-staining analysis of HeLa cells treated with Aceh patchouli oil across a range of concentrations. Each dot plot illustrates the distribution of cell populations across four quadrants: viable cells (Annexin V⁻/PI⁻), early apoptotic cells (Annexin V⁺/PI⁻), late apoptotic cells (Annexin V⁺/PI⁺), and necrotic cells (Annexin V⁻/PI⁺). The data demonstrate a concentration-dependent shift in the cell population toward apoptotic phases, confirming the pro-apoptotic mechanism of patchouli oil. Tabular data include the quantified percentage of cells in each phase across all treatment concentrations and controls, presented as mean ± SD with corresponding statistical analysis, Immunofluorescence images of p-AKT and cleaved-caspase 3 localization in HeLa cells following patchouli oil treatment. - Tables (Tabular Data): Cell viability data (%) at each concentration of patchouli oil. Statistical summary of apoptosis rates (early apoptosis, late apoptosis, necrosis, and viable cells) from flow cytometry analysis. Experimental Context: All experiments were conducted in vitro using HeLa cervical cancer cell lines. Patchouli oil was obtained from Aceh province, Indonesia. Treatments were performed across multiple concentrations with appropriate positive and negative controls. All data are presented as mean ± standard deviation (SD) from at least three independent biological replicates. Intended Use: This dataset supports transparency and reproducibility of the published findings and is made openly available for use by researchers in the fields of natural product pharmacology, oncology, and cell biology.
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Steps to reproduce
All experiments were conducted in vitro using HeLa cervical cancer cell lines maintained in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C with 5% CO₂. Aceh patchouli oil (Pogostemon cablin) was obtained from Aceh province, Indonesia. Stock solutions were prepared in TWEEN-20 and diluted in complete DMEM to working concentrations, with final TWEEN-20 concentration ≤0.01% (v/v) in all groups. Cell Morphology: HeLa cells were seeded in 96-well plates (5 × 10³ cells/well) and incubated for 24 hours. Brightfield images were captured before treatment (baseline) and after treatment with patchouli oil at selected concentrations using an inverted light microscope (10× or 20×). Morphological changes including cell shrinkage, rounding, detachment, and membrane blebbing were documented and saved in TIFF/PNG format (≥300 DPI). CCK-8 Viability Assay: Cells were seeded in 96-well plates (5 × 10³ cells/well) and treated with serial concentrations of patchouli oil for 24–48 hours. CCK-8 reagent (10 µL/well) was added and incubated for 1–4 hours at 37°C. Absorbance was measured at 450 nm. Cell viability (%) was calculated relative to untreated controls and plotted as a dose-response curve. IC₅₀ was determined using GraphPad Prism. Data are expressed as mean ± SD from ≥3 independent replicates. Flow Cytometry (Apoptosis): Cells were seeded in 6-well plates (2–5 × 10⁵ cells/well) and treated with dose-dependent concentrations of patchouli oil. After treatment, cells were collected, washed with cold PBS, and resuspended in Annexin V binding buffer. Annexin V-FITC and Propidium Iodide (PI) staining was performed per manufacturer protocol. A minimum of 10,000 events per sample were acquired on a flow cytometer and analyzed using FlowJo software. Cell populations were quantified as viable (Annexin V⁻/PI⁻), early apoptotic (Annexin V⁺/PI⁻), late apoptotic (Annexin V⁺/PI⁺), and necrotic (Annexin V⁻/PI⁺). Immunofluorescence (p-AKT and Cleaved Caspase-3): Cells grown on glass coverslips were treated with 0.05% patchouli oil, then fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, and blocked with 5% BSA. Cells were incubated overnight at 4°C with primary antibodies against p-AKT and cleaved caspase-3, followed by fluorophore-conjugated secondary antibodies (1 hour, room temperature, dark). Nuclei were counterstained with DAPI. Images were acquired using a fluorescence microscope at consistent exposure settings. Mean Fluorescence Intensity (MFI) was quantified using ImageJ/FIJI from ≥5 fields per condition and presented as bar graphs (mean ± SD). Statistical comparisons were performed using one-way ANOVA with Tukey's post-hoc test. Significance was set at p < 0.05.
Institutions
- University of BrawijayaEast Java, Malang