MOESM1 of Phytochemicals inhibit migration of triple negative breast cancer cells by targeting kinase signaling
Creators: name: Pradip Shahi Thakuri
name: Gupta, Megha
name: Singh, Sunil
name: Ramila Joshi
name: Glasgow, Eric
name: Lekan, Alexander
name: Agarwal, Seema
name: Luker, Gary
name: Tavana, Hossein
Publisher: name: figshare
Year: 2020
Subjects: subject: Biophysics
subject: Biochemistry
subject: Cell Biology
subject: Molecular Biology
subject: Neuroscience
subject: Pharmacology
subject: Biotechnology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer

description: Additional file 1: Table S1. Coordinates of protein kinases in the phospho-kinase dot blot array. Figure S1. (a) Migration of TNBC cells into the cell-excluded gap after 48 hrs without any treatment. (b) Subtypes of TNBC cell lines and their major mutations. Figure S2. Basal-like HCC1937 cells migrated into and occupied the gap almost within 24 hrs. Scale bar is 1 mm. Figure S3. Cytotoxicity analysis TNBC cells treated with (a) fisetin and (b) quercetin. Each data point represents the mean of 8 samples and error bars represent standard error from the mean. (c-e) Representative images of Hs578T show that the spindle-like mesenchymal morphology of cells changes to epithelial cell morphology with fisetin and quercetin treatments. Note that with fisetin treatment of HCC1806 cells and quercetin treatment of HCC1937 cells, a sigmoidal curve could not be fitted to the dose response data. Figure S4. Baseline relative phosphorylation of 43 protein kinases and 2 related signaling proteins in nine TNBC cell lines without any treatments. The hierarchical clustering identified 4 major clusters: Cluster 1 is highlighted with a red box (high baseline activity), Cluster 2 is highlighted with an orange box (high to moderate baseline activity), Cluster 3 is highlighted with a green box (moderate baseline activity), and Cluster 4 is highlighted with blue box (low baseline activity). Figure S5. Fisetin and quercetin treatments are non-cytotoxic to TNBC cells. Viability of nine TNBC cell lines after treatments with (a) 200 µM fisetin and (b) 200 µM quercetin for 6 hours. Figure S6. GSK1059615 treatment dose-dependently downregulated p-WNK1. (a-d) Western blots of p-WNK1 and t-WNK in TNBC cells treated with GSK1059615 for 6 hrs. Figure S7. Combination treatment of TNBC cells with GSK1059615 and WNK463 inhibitors produced an additive effect, suggesting that p-WNK1 is a p-AKT effector. (a) Western blot for single agent and combination treatments for 6 hrs. (b-c) Levels of p-AKT/t-AKT and p-WNK1/t-WNK1 in HCC1806 cells, respectively. ns represents lack of statistically significant difference. Figure S8. CHK2 inhibition promoted migration of different TNBC cells. Images of cell migration (a-c) without any treatment and (d-f) treatments with BML-277. Scale bar is 1 mm. (g) Quantified increased migration of TNBC cells by CHK2 inhibition. Each bar represents a mean of 8 samples, and error bars represent standard error from mean.