Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Overview of peptide sequences and LC 50 values of R-DIM-P-LF11-322 and DIM-LF11-318 [ 30 ].

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Effect of PS-exposure on cell death induced by 5 μM of peptides R-DIM-P-LF11-322 (left) or DIM-LF11-318 (right) in the presence of normal (100%) levels of PS exposed by melanoma cell line A375 (no PSD) or in the presence of reduced levels of PS (1xPSD, 2xPSD and 5xPSD (1xPSD 2,7 μg/10 5 cells) (at least 3 experiments). 5xPSD (13,4 μg/10 5 cells) leads to an approximate reduction of PS-exposure in the outer leaflet (OL) of the plasma membrane of A375 by 40% (at least 2 experiments) . R-DIM-P-LF11-322 in contrast to DIM-LF11-318 shows a significant reduction in killing of melanoma cells in the presence of reduced PS-exposure. PS-exposures was determined by binding to Annexin V-Alexa Fluor 488 (λ ex = 488 nm and λ em = 530 nm). For the measurements serum free media was used to prevent proteolysis of PSD during 30 min of incubation. PSD, PS-decarboxylase PkPSD.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Clinical Proteomics, Membrane, Binding Assay, Incubation

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Distribution of cholesterol in membranes of normal human dermal fibroblasts NHDF (left) and melanoma cells A375 (right). Cholesterol was stained with filipin (green) and nucleus with PI (red). Overlay at the bottom. Pictures are representative results of more than three independent experiments.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Staining

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: A375 cells were treated with 0 mM, 5 mM and 10 mM of cholesterol depletion agent MβCD before peptide incubation. Top: Cholesterol of plasma membrane PM (left) was already efficiently depleted in the presence of 5 mM MβCD (right) as seen by microscopic inspection upon staining with filipin (green). Only intracellular cholesterol remained, presumably located to the Golgi apparatus (GA). Bottom: Upon peptide incubation (10 μM and 20 μM DIM-LF11-322, left; 10 and 20 μM R-DIM-P-LF11-318, right) the PI-uptake of cancer cells, which is proportional to the percentage of killed cells, was measured over four hours. Black represents cell death of A375 in presence of peptides without MβCD treatment, where plasma membrane cholesterol is present. Light gray and gray dashed lines represent cells treated with 5 mM and 10 mM MβCD, respectively. The result indicates a reduction of R-DIM-P-LF11-322 efficacy in presence of PM cholesterol, whereas toxicity of DIM-LF11-318 seems independent on PM cholesterol. Both peptides do not enter cells by clathrin or caveolae mediated endocytosis. Pictures are representative results of three independent experiments. PI-uptake is shown as a mean ± SEM of three independent experiments.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Incubation, Clinical Proteomics, Membrane, Staining

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Melanoma cells (A375) were incubated for two hours with 10 μM fluorescently labeled ((5–6)-FAM-) R-DIM-P-LF11-322. Peptide localization is shown in green, Golgi staining by BODIPY ceramide in red and nuclear staining by NucBlue (blue). Bright field and overlays are illustrated at the bottom. Peptide and Golgi show overlapping staining (yellow) close to the nucleus, suggesting intracellular localization of the peptide to the Golgi apparatus. Peptide (P); Golgi apparatus (GA). Pictures are representative results of six independent experiments.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Incubation, Labeling, Staining

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: A375 with stained cholesterol (filipin, green) were treated with 10 μM R-DIM-P-LF11-322 or DIM-LF11-318 for indicated time periods. Initial localization of cholesterol in PM and GA in untreated melanoma cells changes similarly, however with increased velocity upon treatment with DIM-LF11-318. Both peptides induce consumption of intracellular cholesterol and appearance of cholesterol rich domains in the plasma membrane after 30 min (DIM-LF11-318) or 60–120 min (R-DIM-P-LF11-322). Bottom OL BF: Overlay of bright field (OL BF) and fluorescence (green/filipin, staining of cholesterol; red/PI, staining of DNA of membrane damaged cells) upon 120 min of respective peptide incubation. DIM-LF11-318 causes release of small beads (white arrows), partially composed of cholesterol (green staining). Arrows indicate the plasma membrane (PM), Golgi apparatus (GA) as well as domains (DOM) and beads. Pictures are representative for a series of three experiments.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Staining, Clinical Proteomics, Membrane, Fluorescence, Incubation

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Overlay images of melanoma cells (A375) with stained mitochondria (red, MitoTracker Deep Red) and nucleus (blue, NucBlue) in presence of 10 μM peptide R-DIM-P-LF11-322 (left) or DIM-LF11-318 (right) at different incubation periods of 0, 30, 60 and 120 min. R-DIM-P-LF11-322 causes significant mitochondrial swelling (arrows), whereas DIM-LF11-318 mainly induces cell death indicated by increased uptake of nuclear staining with no significant swelling of mitochondria. Bottom OL BF: Overlay of bright field (OL BF) and respective fluorescence. Arrows indicate membrane debris upon incubation with DIM-LF11-318. Pictures are representative for a series of three experiments.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Staining, Incubation, Fluorescence, Membrane

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Overlay images of melanoma cells (A375) with stained Golgi apparatus (GA), respectively ceramide (red, BODIPYceramide) and nucleus (blue, NucBlue R37606) in presence of 10 μM peptide R-DIM-P-LF11-322 (left) or DIM-LF11-318 (right) at different incubation periods of 0, 30, 60 and 120 min. R-DIM-P-LF11-322 causes distribution of the Golgi marker BODIPY ceramide into the cytosol and slight formation of domains in the plasma membrane (arrows) within 120 min indicating a strong effect on the Golgi apparatus and the distribution of ceramide. DIM-LF11-318 already induces strong staining of the cytosol and plasma membrane after 30 min, indicating even higher stress on the Golgi and formation of punctual domains of ceramide (arrows) in the plasma membrane. Bottom OL BF: Overlay of bright field (OL BF) and respective fluorescence. Arrows indicate membrane debris upon incubation with DIM-LF11-318. Pictures are representative for a series of three experiments.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: Staining, Incubation, Marker, Clinical Proteomics, Membrane, Fluorescence

Journal: PLoS ONE

Article Title: Interaction of two antitumor peptides with membrane lipids – Influence of phosphatidylserine and cholesterol on specificity for melanoma cells

doi: 10.1371/journal.pone.0211187

Figure Lengend Snippet: Summary of characteristics and effects induced by R-DIM-P-LF11-322 and DIM-LF11-318 in model as well as in in vitro systems.

Article Snippet: Normal human dermal fibroblasts (NHDF) and melanoma cells A375 were purchased from PromoCell, Inc. (Heidelberg, Germany) and ATCC (American Type Culture Collection, Manassas, VA, USA), respectively.

Techniques: In Vitro, Sequencing, Zeta Potential Analyzer, Activity Assay, Clinical Proteomics, Membrane, Lysis

Journal: Molecular Systems Biology

Article Title: Integrative multi-omics defines melanoma drug response networks and ARID1A-dependent resistance mechanisms

doi: 10.1038/s44320-025-00183-5

Figure Lengend Snippet: ( A ) Pooled genome-wide CRISPR/Cas9 screens of A375 cells treated with trametinib (MEKi) showing log2 fold change of viability of different knockouts. ( B ) Luminex assay showing the abundance of phosphosites (MEK and ERK) in different A375 genetic contexts after treatment with trametinib. The color shows log 2 fold change in EGFR-stimulated A375 versus serum-free. Data represent biological duplicates, generated using two independent gRNAs targeting the indicated gene. ( C ) Schematic presentation of the method employed to integrate and reconstruct signaling networks from melanoma multi-omics data. Up and down indicate positive and negative weights in the factors. ( D ) Factor weight loadings ( y axis) for the different samples (colors for drug treatment and shapes for genetic conditions). .

Article Snippet: A375 , Synthego , .

Techniques: Genome Wide, CRISPR, Luminex, Generated, Biomarker Discovery

Journal: Molecular Systems Biology

Article Title: Integrative multi-omics defines melanoma drug response networks and ARID1A-dependent resistance mechanisms

doi: 10.1038/s44320-025-00183-5

Figure Lengend Snippet: ( A ) ARID1A and matched parental melanoma A375 cells were purchased from Synthego. Sanger sequencing traces for the edited region is shown. The single base-pair deletion led to a frame-shift mutation in the protein. ( B ) Output from TIDE analysis showing mismatches in the inference window (left panel) and 98% editing efficiency. ( C ) Killing of parental and ARID1A cell lines in the presence of Vermurafenib, Trametinib, combination of both drugs, and control with DMSO. Starting concentration of 30 nM Vermurafenib and 3 nM Trametinib was used (Left panel). The numbers on x-axis indicate the dilution factor (1:3) from the starting concentration. Images of cells at 90 h post treatment with the drugs (Right panel). ( D ) Luminex-based phosphoprotein measurements of indicated phosphosites on A375 cells treated either with Trametinib (10 nM) for indicated times, or with growth factors (TGFb and EGF) for 4 h. Long term indicates treatment with drug at 1 nM for 2 weeks. Log-fold change is calculated from unstimulated/ untreated condition. Decrease in MEK phosphorylation post trametinib treatment is observed between 1 and 6 h.

Article Snippet: A375 , Synthego , .

Techniques: Sequencing, Mutagenesis, Control, Concentration Assay, Luminex, Phospho-proteomics

Journal: Molecular Systems Biology

Article Title: Integrative multi-omics defines melanoma drug response networks and ARID1A-dependent resistance mechanisms

doi: 10.1038/s44320-025-00183-5

Figure Lengend Snippet: ( A ) Nodes ordered ( x axis) by their PageRank centrality ( y axis) in the phuEGO-derived network that are associated with drug-agnostic responses. PageRank centrality reflects the relative influence of each node within the network based on connectivity and neighborhood importance. ( B ) Processes significantly ( y axis) enriched ( x axis) in the drug-agnostic phuEGO-derived network. ( C ) Subset of drug-agnostic phuEGO-derived network highlighting optimized to the 50 most central nodes and their interactions. Semantic similarity indicates the functional similarity between two nodes, as defined by the similarity of their Gene Ontology (GO) terms. The presented mapped protein and transcriptomic factor loadings from MOFA, equivalent to their abundance change in the condition of interest (red increasing after drug treatment and blue decreasing). ( D ) Heatmap demonstrating the changes in proteomics and transcriptomics abundances for negative regulators of the RTK/MAPK pathway (columns) in different drug conditions in ARID1A-WT A375 cells (rows). ( E ) Heatmap demonstrating the changes in functional kinomics measurements in ARID1A-WT cells for multiple kinases (columns) found as regulated or relevant in the drug response network for different drug treatments (rows). For both heatmaps, statistical significance was determined using a two-sided moderated t test (three samples per condition, limma) with Benjamini–Hochberg FDR correction. .

Article Snippet: A375 , Synthego , .

Techniques: Derivative Assay, Functional Assay, Transcriptomics

Journal: Molecular Systems Biology

Article Title: Integrative multi-omics defines melanoma drug response networks and ARID1A-dependent resistance mechanisms

doi: 10.1038/s44320-025-00183-5

Figure Lengend Snippet: ( A ) Kinase activity of selected kinases on ARID1A-KO and WT cells at a basal level and post-combination drug treatment. ( B ) Heatmap demonstrating the changes in proteomic and transcriptomic abundances for selected kinases at a baseline state for ARID1A-KO cells compared to the parental A375 cell line. Statistical significance was determined using a two-sided moderated t test (three samples per condition, limma) with Benjamini–Hochberg FDR correction. ( C ) Expression of EGFR as determined by flow cytometry on the surface of ARID1A-KO or parental A375 ARID1A-WT cell line. ( D ) Scatter plot showing probability distribution from the network propagation from receptors that are upregulated in response to combination therapy (ARID1A-KO, y axis) and receptors that are upregulated in response to ARID1A KO ( x axis). ( E ) Scatter plot showing probability distribution from the network propagation from Ephrin receptors activated following treatment of combination therapy in ARID1A-KO cells. X axis refers to significance (−log10(P)) and the y axis shows the probability distribution of specific nodes. Statistical significance was assessed using an empirical permutation test (10,000 degree-preserving random graph permutations) comparing the stationary probability of each node to its null distribution. ( F ) Network showing how paths from receptors converge on JUN via FYN and PRKD1 following network propagation from EGFR and ROS1. ( G ) Kinase activity assay showing the median kinase activity ( y axis) of selected kinases ( x axis) following combination drug treatment in parental A375 (ARID1A-WT) cell lines (bottom) and ARID1A-KO A375 cell lines (top). ( H ) ARID1A-WT and ARID1A-KO A375 cells expressing Cas9 and mOrange were transduced with empty or EGFR-targeting gRNAs and treated with vemurafenib + trametinib. mOrange loss was monitored over 96 h. ARID1A-KO cells showed slower drug-induced death than WT, a phenotype reversed by EGFR targeting ( n = 4). Statistical significance was assessed using Welch’s two-sample t test (two-sided) for the final time point. .

Article Snippet: A375 , Synthego , .

Techniques: Activity Assay, Expressing, Flow Cytometry, Preserving, Kinase Assay, Transduction

Journal: Molecular Systems Biology

Article Title: Integrative multi-omics defines melanoma drug response networks and ARID1A-dependent resistance mechanisms

doi: 10.1038/s44320-025-00183-5

Figure Lengend Snippet: ( A ) Graph showing the increased connectivity of nodes from both Factor 1 (blue) and Factor 3 (green) compared to Factor 2 (yellow). ( B ) PhuEGO-derived Network underlying changes associated with Factor3. Nodes represent proteins and the edges between them represent known interactions between those proteins. Red indicates networks associated with upregulated processes and blue indicates networks associated with downregulated processes. ( C ) Scatter plots showing the correlation between ARID1A-KO ( y axis) and ARID1A-WT ( x axis) drug response log₂ fold changes in transcriptomic (top row) and proteomic (bottom row) data across different drug conditions (columns). Pearson correlation coefficients (r) and significance values (p-values) were calculated using the Pearson product–moment correlation test, in which significance is assessed using a t-distribution test of the null hypothesis that the true correlation is zero. ( D ) Plot showing the change in activation of TFs MYC (top) and JUN (bottom), based on the relative abundance of their known regulons as calculated using the ColectTRI database. This shows the difference in condition between ARID1A KO A375 cell lines (red) and parental cell lines (blue).

Article Snippet: A375 , Synthego , .

Techniques: Derivative Assay, Activation Assay

Journal: Molecular Systems Biology

Article Title: Integrative multi-omics defines melanoma drug response networks and ARID1A-dependent resistance mechanisms

doi: 10.1038/s44320-025-00183-5

Figure Lengend Snippet: ( A ) Transcription factor activity changes ( x axis) in ARID1A-KO A375 cell line compared to parental ARID1A-WT line. Significance of TF activity was assessed using the t-statistic from a Univariate Linear Model (ULM), where the t value of the regression slope quantifies the likelihood that observed gene expression is explained by the TF’s target-weight model (3 samples per condition, see “Methods”). ( B ) Log-fold changes for regulons ( x axis) of four key transcriptional regulators (CIITA, JUN, RFX5, and TWIST1) that were identified to have significantly differential ( y axis) activity at a basal state for the ARID1A-KO cell line compared to the parental (ARID1A-WT) A375 cell line. Statistical significance was determined using a two-sided moderated t test (three samples per condition, limma) with Benjamini–Hochberg FDR correction. ( C ) Cell surface expression of two MHC class-II antigens (HLA-DQ and HLA-DR) on parental (ARID1A-WT) and ARID1A-KO cell lines. ( D ) Top transcription factors and regulators (e.g., CIITA) with differential activity ( y axis) in ARID1A-mutant versus non-mutant melanoma patients from TCGA. Statistical significance was determined using a univariate linear model (472 samples, see “Methods”). ( E ) Heatmap demonstrating the changes in transcriptomic abundances for selected genes (columns) at a baseline state for either ARID1A-KO compared to parental (ARID1A-WT) A375 cell line (top) or melanoma samples from ARID1A mutant versus WT patients (bottom). .

Article Snippet: A375 , Synthego , .

Techniques: Activity Assay, Gene Expression, Expressing, Mutagenesis

Journal: Frontiers in Oncology

Article Title: Functional Genomic Screening Independently Identifies CUL3 as a Mediator of Vemurafenib Resistance via Src-Rac1 Signaling Axis

doi: 10.3389/fonc.2020.00442

Figure Lengend Snippet: Knockdown of NF1 and CUL3 drive vemurafenib resistance in A375 cells. (A,D) The efficiency of CUL3 (A) and NF1 ( D) knockdown in A375 cells was confirmed by RT-PCR and western blot [* p < 0.05; ** p < 0.01; **** p < 0.0001 (One-way ANOVA followed by Bonferroni's multiple comparisons test)]. (B,E) Sensitivity to vemurafenib was evaluated in short-term (72 h) dose response assay in CUL3 (B) and NF1 (E) A375 knockdown cells. (C,F) Sensitivity to vemurafenib was evaluated in long-term (70 days) growth assay in CUL3 (C) and NF1 (F) A375 knockdown cells [**** p < 0.0001 (Two-way ANOVA followed by Bonferroni's multiple comparisons test)]. n ≥ 3 for each experiment. See for original blots.

Article Snippet: To further filter the genes of interest, expression in A375 was analyzed for each gene, and only genes that had baseline expression in A375 were selected for further study (CCLE, Broad Institute).

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Western Blot, Growth Assay

Journal: Frontiers in Oncology

Article Title: Functional Genomic Screening Independently Identifies CUL3 as a Mediator of Vemurafenib Resistance via Src-Rac1 Signaling Axis

doi: 10.3389/fonc.2020.00442

Figure Lengend Snippet: Resistance to vemurafenib is associated with the reestablishment of MAPK signaling in CUL3 KD cells. (A,B) Western blot of p-ERK1/2 and total ERK1/2 in A375 NF1 KD (A) and CUL3 KD (B) cells treated with DMSO or vemurafenib (3 μM) for 18 h. (C–E) , Expression (C) and quantification (D,E) of phospho-MEK S217 and phospho-MEK S298 measured by western blot in A375 CUL3 KD cells treated with DMSO or vemurafenib (3 μM) for 4 days. * p < 0.05 (Two-way ANOVA followed by Bonferroni's multiple comparisons test), n = 3. See , for original blots.

Article Snippet: To further filter the genes of interest, expression in A375 was analyzed for each gene, and only genes that had baseline expression in A375 were selected for further study (CCLE, Broad Institute).

Techniques: Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: Functional Genomic Screening Independently Identifies CUL3 as a Mediator of Vemurafenib Resistance via Src-Rac1 Signaling Axis

doi: 10.3389/fonc.2020.00442

Figure Lengend Snippet: CUL3 Knockdown cells are resistant to Vemurafenib but sensitive to the combination vemurafenib/saracatinib. (A–C) Expression (A) and quantification of phospho-MEK S298 (B) and phospho-MEK S217 (C) measured by western blot in A375 CUL3 KD cells treated with vemurafenib (3 μM) alone or in combination with saracatinib (2 μM) for 4 days. (D,E) Effect of vemurafenib (3 μM) alone or in combination with saracatinib (2 μM) on the growth of A375 (D) or 451.Lu (E) CUL3 KD cells (10 days treatment). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Two-way ANOVA followed by Bonferroni's multiple comparisons test), n ≥ 3 for each experiment. See for original blots.

Article Snippet: To further filter the genes of interest, expression in A375 was analyzed for each gene, and only genes that had baseline expression in A375 were selected for further study (CCLE, Broad Institute).

Techniques: Knockdown, Expressing, Western Blot

Journal: Frontiers in Oncology

Article Title: Functional Genomic Screening Independently Identifies CUL3 as a Mediator of Vemurafenib Resistance via Src-Rac1 Signaling Axis

doi: 10.3389/fonc.2020.00442

Figure Lengend Snippet: RAC1 activity is increased in CUL3 KD cells and inhibited by the combination vemurafenib/saracatinib. Activity of RAC1 was assessed by pulldown of GTP-RAC1 followed by immunoblotting of RAC1 in (A) A375 and (B) 451.Lu CUL3 KD cells. See , for original blots.

Article Snippet: To further filter the genes of interest, expression in A375 was analyzed for each gene, and only genes that had baseline expression in A375 were selected for further study (CCLE, Broad Institute).

Techniques: Activity Assay, Western Blot

Journal: Frontiers in Oncology

Article Title: Functional Genomic Screening Independently Identifies CUL3 as a Mediator of Vemurafenib Resistance via Src-Rac1 Signaling Axis

doi: 10.3389/fonc.2020.00442

Figure Lengend Snippet: CUL3 KD increases expression of RhoA and Cdc42, while having no effect on RhoC, RAC1, and RhoGDI. (A) Expression and quantification (B) of RhoGTPases (RhoA, RhoC, RAC1, and CDC42) and RhoGDI measured by western blot in A375 CUL3 KD cells. * p < 0.05; **** p < 0.0001 (Two-way ANOVA followed by Bonferroni's multiple comparisons test), n = 3. See for original blots.

Article Snippet: To further filter the genes of interest, expression in A375 was analyzed for each gene, and only genes that had baseline expression in A375 were selected for further study (CCLE, Broad Institute).

Techniques: Expressing, Western Blot

Journal: Frontiers in Oncology

Article Title: Functional Genomic Screening Independently Identifies CUL3 as a Mediator of Vemurafenib Resistance via Src-Rac1 Signaling Axis

doi: 10.3389/fonc.2020.00442

Figure Lengend Snippet: RAC1 KD partially reverses vemurafenib resistance in A375 CUL3 KD cells. (A,B) Sensitivity of RAC1 CUL3 double KD cells to vemurafenib was evaluated in short-term (72 h) dose response assay (1 nM−10 μM). Log IC 60 were calculated and plotted for each cell lines (A) . Log IC 60 and IC 60 values are summarized in (B) . (C) Effect of vemurafenib (3 μM) alone or in combination with saracatinib (2 μM) on the growth of A375 cell line derivatives (5 days treatment). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Two-way ANOVA followed by Bonferroni's multiple comparisons test), n =3.

Article Snippet: To further filter the genes of interest, expression in A375 was analyzed for each gene, and only genes that had baseline expression in A375 were selected for further study (CCLE, Broad Institute).

Techniques:

Journal: Scientific Reports

Article Title: ALDH2 is a novel biomarker and exerts an inhibitory effect on melanoma

doi: 10.1038/s41598-024-54084-y

Figure Lengend Snippet: The mRNA expression of ALDH2, ADH1B, ALDH3A2, DPT, EPHX2, and GATM was measured in melanoma cells (A375/SK-MEL-2 cells) and human normal melanin cell line (PIG1 cells) by quantitative real-time polymerase chain reaction (qRT-PCR), ** P < 0.01, *** P < 0.001 versus PIG1 cells.

Article Snippet: Human normal melanin cell line, PIG1 (American type culture collection, Manzas, Virginia, USA), and malignant melanoma cells, A375 and SK-MEL-2 cells (iCell Bioscience Inc, Shanghai, China) were maintained in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum at 37° C with 5% CO 2 .

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: ALDH2 is a novel biomarker and exerts an inhibitory effect on melanoma

doi: 10.1038/s41598-024-54084-y

Figure Lengend Snippet: ALDH2 inhibits viability, proliferation, migration, and invasion of melanoma cells. ( A ) Transfection efficiency was detected by qRT-PCR. ( B ) The viability of A375/SK-MEL-2 cells was measured by Cell counting Kit-8 assay. ( C ) The proliferation was detected by 5-Ethynyl-2′-deoxyuridine assay. ( D ) The migration of A375/SK-MEL-2 cells was measured by wound healing assay. ( E ) The invasion of A375/SK-MEL-2 cells was measured by Transwell assay. *** P < 0.001 versus pcDNA3.1-NC group.

Article Snippet: Human normal melanin cell line, PIG1 (American type culture collection, Manzas, Virginia, USA), and malignant melanoma cells, A375 and SK-MEL-2 cells (iCell Bioscience Inc, Shanghai, China) were maintained in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum at 37° C with 5% CO 2 .

Techniques: Migration, Transfection, Quantitative RT-PCR, Cell Counting, Wound Healing Assay, Transwell Assay