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. 2022 Aug 24;13(8):730. doi: 10.1038/s41419-022-05177-x

Fig. 3. Changes in the activity of the PEPCK-M pathway regulates entosis in the absence of glucose.

Fig. 3

A Percentage of entotic cells observed in wild-type MCF7 cells after 72 h of growth in 0 mM glucose media in the presence of iPEPCK-2 (5 µM) or vehicle (DMSO) (Two-tailed t-test; n = 5). B Percentage of entotic cells observed in wild-type MCF7 cells after 48 h of growth in 25 mM glucose media in the presence of iPEPCK-2 (5 µM) or vehicle (DMSO) (Two-tailed t-test; n = 3). C Percentage of apoptotic+necrotic cells wild-type MCF7 cells detected after 72 h of growth in 0 mM glucose media in the presence of iPEPCK-2 (5 µM), and Y27632 (10 µM) or vehicle (DMSO). Cells were analyzed by flow cytometry using Annexin V-APC and SYTOX™ Green Nucleic Acid Stain Statistical analysis queried Y27632 versus DMSO in the presence of iPEPCK-2 (Two-tailed t-test; n = 6). D Percentage of entotic cells observed in wild-type MCF7 cells and in MCF7 cells overexpressing PEPCK-M (L-PCK2) at the 72 h time point of growth in 0 mM glucose media (Two-tailed t-test; n = 4). E Percentage of apoptotic+necrotic cells wild-type MCF7 and in MCF7 cells overexpressing PEPCK-M (L-PCK2) cells detected after 72 h of growth in 0 mM glucose media in the presence of Y27632 (10 µM) or vehicle (DMSO). Cells were analyzed by flow cytometry using Annexin V-APC and SYTOX™ Green Nucleic Acid Stain Statistical analysis queried MCF7-L versus MCF7 WT (#One-way ANOVA with Newman–Keuls post-hoc test; n = 8) or Y27632 versus DMSO (*Two-tailed t-test; n = 8). F Percentage of entotic cells observed in wild-type MCF7 cells detected after 24 h of growth in 25 mM glucose media with taxol (1 µM) in the presence of iPEPCK-2 (5 µM) or vehicle (DMSO) (Two-tailed t-test; n = 4). G Percentage of entosis in wild-type MCF7 cells grown in 25 mM glucose media in the presence of thapsigargin (100 nM; 24 h) and amino-acid limitation media (DMEM w/o serine, glycine, arginine and leucine; 48 h). Additionally, cells were treated with iPEPCK-2 (5 µM) or vehicle (DMSO). Statistical analysis queried iPEPCK-2 versus DMSO for each independent treatment; Thapsi and AA Depr (Two-tailed t-test; n = 8).