free cholesterol e kit  (FUJIFILM)

Journal: Frontiers in Molecular Biosciences

Article Title: SOX2 induces LPCAT1 expression to promote cholesterol metabolic reprogramming-mediated invasion and metastasis in osteosarcoma

doi: 10.3389/fmolb.2025.1679244

Figure Lengend Snippet: RNA sequencing transcriptomic analysis. (A,B) Volcano plot and heatmap showing DEGs in MG63 cells after LPCAT1 overexpression. (C,D) Volcano plot and heatmap displaying DEGs in U2OS cells following LPCAT1 knockdown. (E,F) GO enrichment analysis revealing LPCAT1-regulated cholesterol biosynthesis processes in both MG63 and U2OS cells. (G) Reactome pathway analysis visualizing LPCAT1-affected genes in osteosarcoma cells.

Article Snippet: Free cholesterol (FC, E-BC-K004-M) and total cholesterol (TC, E-BC-K109-M) levels in cells (5 × 10 6 ) or tissues (50 mg) were measured using commercial kits (Elabscience, China).

Techniques: RNA Sequencing, Over Expression, Knockdown

Journal: Frontiers in Molecular Biosciences

Article Title: SOX2 induces LPCAT1 expression to promote cholesterol metabolic reprogramming-mediated invasion and metastasis in osteosarcoma

doi: 10.3389/fmolb.2025.1679244

Figure Lengend Snippet: Role of LPCAT1 in cholesterol metabolic reprogramming of osteosarcoma cells. (A,B) Cholesterol content measurement in osteosarcoma cells after LPCAT1 modulation using commercial assay kits. (C,D) Western blot and qRT-PCR analysis of cholesterol metabolism-related proteins following LPCAT1 overexpression or knockdown. Data are representative of three independent experiments (mean ± SD). ** P < 0.01, *** P < 0.001 vs. NC.

Article Snippet: Free cholesterol (FC, E-BC-K004-M) and total cholesterol (TC, E-BC-K109-M) levels in cells (5 × 10 6 ) or tissues (50 mg) were measured using commercial kits (Elabscience, China).

Techniques: Western Blot, Quantitative RT-PCR, Over Expression, Knockdown

Journal: Frontiers in Molecular Biosciences

Article Title: SOX2 induces LPCAT1 expression to promote cholesterol metabolic reprogramming-mediated invasion and metastasis in osteosarcoma

doi: 10.3389/fmolb.2025.1679244

Figure Lengend Snippet: In vitro validation of SOX2/LPCAT1 axis in regulating osteosarcoma cell biological behaviors and cholesterol metabolism. MG63 cells were co-transfected with oe-LPCAT1 and/or shSOX2, while U2OS cells were co-transfected with shLPCAT1 and/or SOX2. (A,B) Cell viability was determined by CCK-8 assay. (C,D) Migration ability was assessed by wound healing assay. (E,F) Invasion capacity was evaluated by Transwell assay. (G,H) Intracellular cholesterol level measurement. (I,J) Western blot and qRT-PCR analysis of metastasis- and cholesterol metabolism-related proteins. Data are representative of three independent experiments (mean ± SD). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. NC+shNC, shLPCAT1+NC or LPCAT1+shNC, SOX2+shNC or shSOX2+NC.

Article Snippet: Free cholesterol (FC, E-BC-K004-M) and total cholesterol (TC, E-BC-K109-M) levels in cells (5 × 10 6 ) or tissues (50 mg) were measured using commercial kits (Elabscience, China).

Techniques: In Vitro, Biomarker Discovery, Transfection, CCK-8 Assay, Migration, Wound Healing Assay, Transwell Assay, Western Blot, Quantitative RT-PCR

Journal: Frontiers in Molecular Biosciences

Article Title: SOX2 induces LPCAT1 expression to promote cholesterol metabolic reprogramming-mediated invasion and metastasis in osteosarcoma

doi: 10.3389/fmolb.2025.1679244

Figure Lengend Snippet: In vivo validation of SOX2/LPCAT1 axis in osteosarcoma progression and cholesterol metabolism. For (A–D) , MG63 cells were transfected with oe-LPCAT1 and/or shSOX2, and then subcutaneously injected into nude mice to construct a xenograft model. (A) Representative images of xenograft tumors. (B) Tumor volume. (C) Cholesterol content measurement in tumor tissues. (D,E) Western blot analysis of metastasis- and cholesterol metabolism-related proteins in xenograft tumors. (F) MG63 cells transfected with oe-LPCAT1 and/or shSOX2 were injected into the tail vein of nude mice to construct a lung metastasis model. The number of pulmonary nodules was counted. Data are representative of three independent experiments (mean ± SD). ** P < 0.01, *** P < 0.001 vs. NC+shNC, LPCAT1+shNC, and shSOX2+NC.

Article Snippet: Free cholesterol (FC, E-BC-K004-M) and total cholesterol (TC, E-BC-K109-M) levels in cells (5 × 10 6 ) or tissues (50 mg) were measured using commercial kits (Elabscience, China).

Techniques: In Vivo, Biomarker Discovery, Transfection, Injection, Construct, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: APP family is a regulator of endo-lysosomal membrane vulnerability

doi: 10.1016/j.jbc.2025.110774

Figure Lengend Snippet: Comprehensive lipid profiles in the membrane fraction of APP / APLP2 dKO MEF cells and WT controls. A, the same number of APP/APLP2 dKO or WT MEF cells was subjected to subcellular fractionation, and total cholesterol levels in their membrane fractions were measured using the total cholesterol colorimetric assay kit. Total cholesterol levels significantly increased in APP/APLP2 dKO MEF cells compared with WT controls. N = 3. Unpaired t test, ∗∗ p < 0.01. B, the membrane fraction of APP/APLP2 dKO MEF cells or WT controls was subjected to LC–MS/MS lipidomic analysis, and the signal area (a.u.) was measured for various lipid species (N = 4 independent experiments). The analysis recapitulated the significantly increased total cholesterol level, which is based on the sum of all the detected cholesterol species, in APP/APLP2 dKO compared with WT MEF cells. Unpaired t test, ∗ p < 0.05. C, the heatmap and bar graphs were generated from log-transformed area and the exact area data, respectively, showing differences in individual cholesterol species between APP/APLP2 dKO MEF cells and WT controls. Dots in bar graphs correspond to the values in individual experiments, and bar graphs show means. Mann–Whitney U test, ∗ p < 0.05. D, similarly, the heatmap and bar graphs are shown for other lipid species, where the areas of all the lipid species are combined. Mann–Whitney U test, ∗∗ p < 0.01, ∗ p < 0.05. APLP, amyloid precursor–like protein; APP, amyloid precursor protein; dKO, double KO; MEF, mouse embryonic fibroblast.

Article Snippet: To measure total cholesterol levels, including free cholesterol and cholesterol esters, the total cholesterol colorimetric assay kit (Elabscience) was used according to the manufacturer’s protocol.

Techniques: Membrane, Fractionation, Colorimetric Assay, Liquid Chromatography with Mass Spectroscopy, Generated, Transformation Assay, MANN-WHITNEY