mg63 os cells  (iCell Bioscience Inc)

Journal: bioRxiv

Article Title: Proteome-wide multi-omics profiling of osteosarcoma transcription factor networks

doi: 10.64898/2026.03.29.714917

Figure Lengend Snippet: A) Schema of the concatenated tandem array of the consensus transcription factor response elements (catTFREs) workflow utilising two plasmids inserts C1 (designed with approximately 110 TF binding sites (TFBS) in duplicates with linkers) and C4 (reduced TFBS). Detailed TFBS are provided in Table S1A and B. Inserts were linearized by restriction enzyme digestion and biotinylated by Klenow enzyme. Biotinylated C1 and C4 were immobilized on streptavidin coated magnetic beads and incubated with nuclear extracts (NE) for detection of DNA binding proteins by mass spectrometry (MS). B) MS spectral counts of proteins enriched from NEs or by TFRE C1 and C4 pulldown. Four replicates with total count of proteins (left) and TFs only (right) from HOS (purple), U2OS (red) and MG63 (blue). All source data are provided in Table S1C-F. C) Heatmap representing the Pearson correlation of spectral counts by samples from HOS, U2OS and MG63. Scale represents low correlation in white to self-correlations as red-brown. D) Clustering of proteins by Weighted Gene Co-expression Network Analysis from MS spectral counts of NE and C1/C4 in HOS, MG63 and U2OS. Clusters I to X with the number of proteins represented by Pearson correlation across all conditions. Proteins are listed in Table S1G.

Article Snippet: Human osteosarcoma HOS, U2OS, and MG63 cell lines (CRL-1543, HTB-96 and CRL-1427, .) were purchased from the American Type Culture Collection (ATCC).

Techniques: Binding Assay, Magnetic Beads, Incubation, DNA Binding Assay, Mass Spectrometry, Expressing

Journal: bioRxiv

Article Title: Proteome-wide multi-omics profiling of osteosarcoma transcription factor networks

doi: 10.64898/2026.03.29.714917

Figure Lengend Snippet: A) Heat map of differential protein analysis between cell lines, C1, C4 and NE comparisons, defining clusters 1-6 (n = 4). B) Intensity levels of proteins JUNB and RUNX2 above; and TEAD1 and NFIB below, in NE and the catTFREs C1 and C4, for cell lines MG63, U2OS and HOS (n = 4). C, D, and E) Volcano plots of differential protein expression (DEP) analysis between C1 and NE in MG63 (in C), U2OS (in D), and HOS (in E), abs(log2FoldChange) > 1 and a p-value < 0.05. Significant proteins detected are coloured in blue (NE) and red (C1) with a selection of key proteins highlighted. All source data are provided in Table S2.

Article Snippet: Human osteosarcoma HOS, U2OS, and MG63 cell lines (CRL-1543, HTB-96 and CRL-1427, .) were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, Selection

Journal: bioRxiv

Article Title: Proteome-wide multi-omics profiling of osteosarcoma transcription factor networks

doi: 10.64898/2026.03.29.714917

Figure Lengend Snippet: A) Volcano plots of the differential protein expression (DEPs) between catTFRE C1 for HOS (red) and MG63 (blue) cell lines. B) Volcano plots of the differential protein expression (DEPs) between catTFRE C1 for U2OS (red) and MG63 (blue) cell lines. C) Integration of the differential protein analysis for C1 from the HOS/MG63 and U2OS/MG63 comparison by log2Foldchanges and p-adjust values. Significant genes are clustered by log2FoldChange, highlighted by −log10(pValue HOS/MG63) and sized by −log10(pValue U2OS/MG63). D) Top 200 of the highest differential edge degrees between HOS/MG63, filtering based on the differential protein enrichment in A), plotted by Cytoscape. The edges in HOS and MG63 were highlighted in purple and green, respectively. E) Top 200 of the highest differential edge degrees between U2OS/MG63, filtering based on the differential protein enrichment in B). The edges in U2OS and MG63 were highlighted in purple and green, respectively. All source data are provided in Table S3.

Article Snippet: Human osteosarcoma HOS, U2OS, and MG63 cell lines (CRL-1543, HTB-96 and CRL-1427, .) were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, Comparison, Protein Enrichment

Journal: bioRxiv

Article Title: Proteome-wide multi-omics profiling of osteosarcoma transcription factor networks

doi: 10.64898/2026.03.29.714917

Figure Lengend Snippet: A) and B) Differential gene expression (DEGs) in the comparison of U2OS/MG63 and HOS/MG63. Statistically significant higher expression of HOS and U2OS genes are highlighted in red, while blue indicates higher expression of MG63. D) and E) Integration of the logFoldChange of DEPs and DEGs of U2OS/MG63 (in A) and HOS/MG63 (in B), with |log2FoldChange catTFREs C1| > 1 and |log2FoldChange RNA-seq| > 1. Four clusters are indicated by colours red, blue, purple, and yellow. Each cluster has a different number of proteins and key genes annotated. E) A Venn diagram of integrated datasets of U2OS and HOS for catTFRE up and RNA-seq up. All source data are provided in Table S4.

Article Snippet: Human osteosarcoma HOS, U2OS, and MG63 cell lines (CRL-1543, HTB-96 and CRL-1427, .) were purchased from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Comparison, Expressing, RNA Sequencing

Journal: bioRxiv

Article Title: Proteome-wide multi-omics profiling of osteosarcoma transcription factor networks

doi: 10.64898/2026.03.29.714917

Figure Lengend Snippet: A) Heatmap visualizing the correlation of catTFREs C1 MS , RNA-seq , and consensus peaks at TSS (ATAC-seq DARs) that are highly correlated. All genes/proteins were filtered based on the Pearson correlation between each pair of omics, where at least one pair had a correlation coefficient greater than 0.5. Four clusters are annotated where A is specific to MG63, B specific to HOS/U2OS, C specific to HOS and D specific to U2OS. B) Overview of high confidence of the predicted protein–protein interactions using the StringDb database. The cluster represents the protein and its interaction greater than 0.4 from the HOS/U2OS-shared cluster (Cluster B). C) Heatmap visualizing the normalized gene expression RPKM of all the TFs enriched in footprinting analysis in MG63, HOS and U2OS that were significant in catTFRE C1 comparison in , and . Four replicates of RNA-seq per cell line. D) Ranking of TF footprinting from whole-genome consensus peaks in MG63 (left), HOS (middle) and U2OS (right) using chromVAR. TFs are coloured in blue (not significant), magenta (significant), and grey (not detected) for catTFREs C1 differential comparison from , and and integrated with RNA-seq, MG63/HOS or MG63/U2OS for each cell line. All source data are provided in Table S5.

Article Snippet: Human osteosarcoma HOS, U2OS, and MG63 cell lines (CRL-1543, HTB-96 and CRL-1427, .) were purchased from the American Type Culture Collection (ATCC).

Techniques: RNA Sequencing, Protein-Protein interactions, Gene Expression, Footprinting, Comparison

Journal: Frontiers in Pharmacology

Article Title: Shikonin suppresses proliferation of osteosarcoma cells by inducing ferroptosis through promoting Nrf2 ubiquitination and inhibiting the xCT/GPX4 regulatory axis

doi: 10.3389/fphar.2024.1490759

Figure Lengend Snippet: Shikonin inhibited cell proliferation of MG63 and 143B cells. (A) The molecular and 3D structure of shikonin. (B–C) MG63 and 143B cells were cultured at different concentrations of shikonin (0, 0.25, 0.5, 1, 2, 4, 6, 8 and 10 uM) for 24, 48 h, and then CCK-8 assay was performed to detect the cell proliferation. (D) Giemsa staining was used to observe the effect of shikonin on cell morphology. (E–F) Colony formation and (G–I) EdU Staining were used to observe the impact of shikonin on cell proliferation. (J, K) Flow cytometry was used to analyze the effects of shikonin on the cell cycle when cells were treated with or without shikonin for 24 h (L, M) Western blot was used to detect the expression of proliferation protein PCNA. * p < 0.05, ** p < 0.01, *** p < 0.001. The data were presented as mean ± SD, and analyzed using one-way ANOVA following Tukey’s t-test (n = 3).

Article Snippet: The OS cell lines MG63 and143B were originally obtained from the American Type Culture Collection (ATCC) and were maintained at 37°C in a humidified atmosphere consisting of 5% CO 2 .

Techniques: Cell Culture, CCK-8 Assay, Staining, Flow Cytometry, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Shikonin suppresses proliferation of osteosarcoma cells by inducing ferroptosis through promoting Nrf2 ubiquitination and inhibiting the xCT/GPX4 regulatory axis

doi: 10.3389/fphar.2024.1490759

Figure Lengend Snippet: Shikonin triggered the ferroptosis in OS cells. (A, B) The MG63 and 143B cells were co-treated with shikonin and Fer-1 after 24 h, and the cell viabilities were examined by CCK-8 assay. (C–F) The ROS, (G–J) lipid peroxidation and (K–N) Fe 2+ contents were performed by Flow cytometry, followed by quantitative analysis. (O, P) Western blot was used to detect the expression of protein xCT and GPX4, and quantification was analyzed. The levels of MDA (Q) and GSH/GSSG (R) were determined in the shikonin treated OS cells. * p < 0.05, ** p < 0.01, *** p < 0.001. The data were presented as mean ± SD, and analyzed using one-way ANOVA following Tukey’s t-test (n = 3).

Article Snippet: The OS cell lines MG63 and143B were originally obtained from the American Type Culture Collection (ATCC) and were maintained at 37°C in a humidified atmosphere consisting of 5% CO 2 .

Techniques: CCK-8 Assay, Flow Cytometry, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Shikonin suppresses proliferation of osteosarcoma cells by inducing ferroptosis through promoting Nrf2 ubiquitination and inhibiting the xCT/GPX4 regulatory axis

doi: 10.3389/fphar.2024.1490759

Figure Lengend Snippet: Shikonin induced mitochondria damage in OS cells. (A, B) The mitochondrial morphology of MG63 and 143B cells treated with or without shikonin after 24 h was observed by transmission electron microscopy (scale bar = 1 um). The red arrows represented mitochondrial changes typical of ferroptosis. (C, D) Mitochondrial Fe 2+ and (E, F) mitochondrial ROS contents was observed by using Laser Scanning Confocal Microscopy (LSCM) after the intervention of co-treated with shikonin and Fer-1 after 24 h in MG63 and 143B cells (scale bar = 100 um).

Article Snippet: The OS cell lines MG63 and143B were originally obtained from the American Type Culture Collection (ATCC) and were maintained at 37°C in a humidified atmosphere consisting of 5% CO 2 .

Techniques: Transmission Assay, Electron Microscopy, Confocal Microscopy

Journal: Frontiers in Pharmacology

Article Title: Shikonin suppresses proliferation of osteosarcoma cells by inducing ferroptosis through promoting Nrf2 ubiquitination and inhibiting the xCT/GPX4 regulatory axis

doi: 10.3389/fphar.2024.1490759

Figure Lengend Snippet: Shikonin physically interacted with Nrf2 and promoted Nrf2 degradation in OS cells. (A, B) Western blot was used to detect the expression of Nrf2 and keap1 in shikonin-treated MG63 and 143B cells, and quantification was analyzed. (C) Nrf2 mRNA levels in the control or shikonin (1.5 uM)-treated MG63 and 143B cells for 24 h. (D) The interaction between shikonin with Nrf2 was predicted by molecular docking. (E) Co-immunoprecipitation and Western blot assay of Nrf2 protein after shikonin and MG132 (2.5 uM) co-treatment 24 h in MG63 and 143B cells. (F, G) Western blot was used to observe the expression of the Nrf2 protein after the intervention of 1.5 uM shikonin cotreated with or without MG132 (2.5 uM) and CHX (35 uM) for 24 h, and quantification was analyzed. * p < 0.05, ** p < 0.01, *** p < 0.001. The data were presented as mean ± SD, and analyzed using one-way ANOVA following Tukey’s t-test (n = 3).

Article Snippet: The OS cell lines MG63 and143B were originally obtained from the American Type Culture Collection (ATCC) and were maintained at 37°C in a humidified atmosphere consisting of 5% CO 2 .

Techniques: Western Blot, Expressing, Control, Immunoprecipitation

Journal: Frontiers in Pharmacology

Article Title: Shikonin suppresses proliferation of osteosarcoma cells by inducing ferroptosis through promoting Nrf2 ubiquitination and inhibiting the xCT/GPX4 regulatory axis

doi: 10.3389/fphar.2024.1490759

Figure Lengend Snippet: Overexpression of Nrf2 reversed the Shikonin-induced ferroptosis in OS cells. (A–D) Western blot was used to detect the expression of protein level of Nrf2, xCT and GPX4 in the shikonin treated Nrf2 overexpressing MG63 and 143B cells. (E–G) Flow cytometry was used to observe the total lipid peroxidation levels after the intervention of shikonin (1.5 uM) for 24 h in Nrf2 overexpressing MG63 and 143B cells. * p < 0.05, ** p < 0.01, *** p < 0.001. The data were presented as mean ± SD, and analyzed using one-way ANOVA following Tukey’s t-test (n = 3).

Article Snippet: The OS cell lines MG63 and143B were originally obtained from the American Type Culture Collection (ATCC) and were maintained at 37°C in a humidified atmosphere consisting of 5% CO 2 .

Techniques: Over Expression, Western Blot, Expressing, Flow Cytometry