Journal: Breast Cancer : Targets and Therapy

Article Title: Identification and Validation of a Novel Theranostic Target in Triple Negative Breast Cancer with Transcriptomics and Protein Analyses

doi: 10.2147/BCTT.S568001

Figure Lengend Snippet: Validation of LY6E expression in TNBC cell lines and xenograft tumors. ( A ) Western blot analysis of whole-cell lysates from HMEC and TNBC cell lines. LY6E was detected in MDA-MB-468, HCC-1143, and HCC-1937, and showed weak expression in HMEC. ( B ) Flow cytometry analysis of MDA-MB-468 cells confirmed membrane localization of LY6E (red) compared with isotype control (gray). ( C ) Immunofluorescence confocal microscopy of MDA-MB-468 cells showing LY6E staining (green) localized to the cell membrane. Nuclei were counterstained with DAPI (blue). ( D ) Western blot analysis of tumors derived from MDA-MB-468 xenografts demonstrating consistent LY6E expression.

Article Snippet: MDA-MB-231, MDA-MB-468, HCC-1143, and HCC-1937 were selected as representative TNBC cell lines based on transcriptomics results from human cancer cell lines., HMEC, MDA-MB-231, and MDA-MB-468 cells were purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Biomarker Discovery, Expressing, Western Blot, Flow Cytometry, Membrane, Control, Immunofluorescence, Confocal Microscopy, Staining, Derivative Assay

Journal: Scientific Reports

Article Title: The novel amino-artemisinin derivative WHN-11 disrupts mitochondria and protein homeostasis, and induces autophagy and apoptosis in cancer cells

doi: 10.1038/s41598-025-05284-7

Figure Lengend Snippet: WHN-11 disrupts mitochondrial function. ( a ) Analysis of Bcl-xL expression in treated HCC1937 cells overnight. Data indicate the mean fluorescence intensity ± SEM (n = 3), where significance levels are indicated as * p < 0.05 using a two-tailed student’s t-test. ( b ) Cellular ATP levels quantified in treated cells. Data shown are averages ± SEM (n = 3), where significance levels are indicated as * p < 0.05, ** p < 0.01 and *** p < 0.001 using a two-tailed student’s t-test. ( c ) Mitochondrial membrane potential (ΔΨm) measured by JC-1 staining in treated HCC1937 cells. Data shown are averages ± SEM (n = 3) of the relative percentage ratio of red to green fluorescence, where significance levels are indicated as ** p < 0.01, *** p < 0.001 and ns–not significant using a two-tailed student’s t-test. ( d ) Confocal analysis of the mitochondrial morphology indicated by TRAP1 and Bcl-xL staining in treated HCC1937 cells. Representative images shown (n = 2).

Article Snippet: ATP levels in treated HCC1937 cells was assessed using the ATP Bioluminescence Assay Kit CLS II (Roche, 11–699-695–001).

Techniques: Expressing, Fluorescence, Two Tailed Test, Membrane, Staining

Journal: Molecular & Cellular Proteomics

Article Title: Quantification of the N-glycosylated Secretome by Super-SILAC During Breast Cancer Progression and in Human Blood Samples

doi: 10.1074/mcp.m112.023614

Figure Lengend Snippet: FIG. 1. Overview of the experimental workflow. A, The secretome was collected as conditioned medium from cell lines in culture. HCC1143, HCC1937 and HCC2218 were also cultured under SILAC conditions to generate a super-SILAC mix as an internal standard. B, N-glycosylated peptides were enriched using N-glyco FASP. Proteins were first digested to peptides and N-glycosylated peptides were then captured using lectins. Finally, N-glycosylated peptides were released by deglycosylation using PNGase F and the deglycosylated peptides were analyzed by LC-MS.

Article Snippet: HCC202, and HCC2218 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA); HCC1143, HCC1937, and HCC1599 cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ); MCF-10a and MDA-MB-453 cells were kindly provided by Axel Ulrich (Max-Planck Institute of Biochemistry, Martinsried, Germany).

Techniques: Cell Culture, Multiplex sample analysis, Liquid Chromatography with Mass Spectroscopy

Journal: Molecular & Cellular Proteomics

Article Title: Quantification of the N-glycosylated Secretome by Super-SILAC During Breast Cancer Progression and in Human Blood Samples

doi: 10.1074/mcp.m112.023614

Figure Lengend Snippet: FIG. 3. Effect of normalization using the super-SILAC internal standard. A, Correlation of intensities of light N-glycosylated peptides intensities between different replicates and cell lines (light channel of SILAC measurements only). Samples are ordered horizontally and vertically according to replicates and cell line (from HMEpC1 until MDA-MB-453). B, Correlation of intensities of heavy isotope labeled N-glycosylated peptides (super-SILAC internal standard) intensities between different replicates and cell lines. C, Correlation of N-glycosylated peptide ratios between different replicates and cell lines after normalization using the super-SILAC internal standard. Correlations between replicates (“green blocks” near the diagonal) and cell lines from the same cancer stages are increased, whereas differences with other cell lines are augmented. D, Correlation between replicates from each cell line at the N-glycosylated peptide level (from panel A and C) and protein level (based on nonglycosylated and glycosylated peptides) before and after normalization using the super-SILAC internal standard. Pearson’s r was calculated between all replicates from each cell line. Two aberrant measurements (one from HMEpC2 and one from HCC1937) were omitted.

Article Snippet: HCC202, and HCC2218 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA); HCC1143, HCC1937, and HCC1599 cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ); MCF-10a and MDA-MB-453 cells were kindly provided by Axel Ulrich (Max-Planck Institute of Biochemistry, Martinsried, Germany).

Techniques: Multiplex sample analysis, Labeling