Journal: Matrix Biology Plus
Article Title: Profiling of collagen and extracellular matrix deposition from cell culture using in vitro ExtraCellular matrix mass spectrometry imaging (ivECM-MSI)
doi: 10.1016/j.mbplus.2024.100161
Figure Lengend Snippet: Deposition of collagen in H1299 lung carcinoma cells treated with DMSO and NS1643 Kv11.1 potassium channel activator. A) Identification and quantification of cellular senescence by flow cytometry of representative DNA content in H1299 cells before and after NS1643 treatment (50 µM for 24 h). Protein level is p21 in control (C) vs NS1643 treated cells. B) Percent change of cell accumulation for G0/G1, S and G2/M phases of the cell cycle (n = 3 technical replicates (n = well); *=p < 0.05). C) Example western blots showing the expression level of Kv11.1 and senescent marker p21 cip1/waf1 in H1299 cells treated with DMSO and NS1643 (50 µM/24 h; n = 3; *=p < 0.05). No change of Kv11.1 protein was observed after treatment (control). D) Senescence-associated β-galactosidase (SA-β-gal) staining. Representative images of H1299 cells treated with (DMSO, left) or NS1643 (50uM/24hr, right) as indicated. Cells were treated with the Kv11.1 activator NS1643 (50 μM/48 hr) to induce cellular senescence. Histogram of average well peak intensities for the putative [M+H] + peptide for E) fibril forming collagen α1(I) and F) fibril-associated collagen α1(XIV) for both slides seeded at 2E5 cells. The center of each data point is the mean and error bars equate to the standard deviation. After one-way ANOVA analysis and Tukey multiple comparisons test, only significant Tukey p-values ≤ 0.05 are displayed. (ox) = methionine oxidation; ppm- parts per million match by mass accuracy. ivECM-MSI measurements were on two four chamber well slides three separate wells.
Article Snippet: All antibodies were purchased from Cell Signaling Technologies, Inc (p21cip1/waf1 Cat 2947; Histone H3 Cat# 4499).
Techniques: Flow Cytometry, Control, Western Blot, Expressing, Marker, Staining, Standard Deviation
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![Deposition of collagen in H1299 lung carcinoma cells treated with DMSO and NS1643 Kv11.1 potassium channel activator. A) Identification and quantification of cellular senescence by flow cytometry of representative DNA content in H1299 cells before and after NS1643 treatment (50 µM for 24 h). Protein level is p21 in control (C) vs NS1643 treated cells. B) Percent change of cell accumulation for G0/G1, S and G2/M phases of the cell cycle (n = 3 technical replicates (n = well); *=p < 0.05). C) Example western blots showing the expression level of Kv11.1 and senescent marker p21 <t>cip1/waf1</t> in H1299 cells treated with DMSO and NS1643 (50 µM/24 h; n = 3; *=p < 0.05). No change of Kv11.1 protein was observed after treatment (control). D) Senescence-associated β-galactosidase (SA-β-gal) staining. Representative images of H1299 cells treated with (DMSO, left) or NS1643 (50uM/24hr, right) as indicated. Cells were treated with the Kv11.1 activator NS1643 (50 μM/48 hr) to induce cellular senescence. Histogram of average well peak intensities for the putative [M+H] + peptide for E) fibril forming collagen α1(I) and F) fibril-associated collagen α1(XIV) for both slides seeded at 2E5 cells. The center of each data point is the mean and error bars equate to the standard deviation. After one-way ANOVA analysis and Tukey multiple comparisons test, only significant Tukey p-values ≤ 0.05 are displayed. (ox) = methionine oxidation; ppm- parts per million match by mass accuracy. ivECM-MSI measurements were on two four chamber well slides three separate wells.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_2733/pmc11492733/pmc11492733__gr4.jpg)