Figure 4. ROS prevents exocyst binding... | Rockefeller University Press

Figure 4.

$ROS prevents exocyst binding to the membrane. (A) A diagram describing the MCF7 cell model used to examine the impact of a 30-min ROS exposure on the membrane proteome. (B) A volcano plot examining the changes in membrane proteome caused by ROS exposure (FDR < 0.05). (C) A heat map depicting the expression changes of the top 100 most differentially regulated proteins in membranes isolated from ROS-exposed cells (FDR < 0.05). (D) Protein abundances for several common membrane proteins in control and H2O2-treated cells. (E) Membrane abundances for exocyst complex components in control and H2O2-treated MCF7 cells (FDR < 0.05). (F) A heat map showing the top 100 most differentially regulated proteins in membrane fraction from 3T3 cells and 3T3 treated with H2O2. (G) A heat map representing proteomic-based measurements of the detectable subunits of the exocyst complex from membrane fractions isolated from Drosophila eggs. (H) A heat map representing proteomic-based measurements of the detectable subunits of the exocyst complex from membrane fractions isolated from NIH3T3 cells. (I) A heat map representing proteomic-based measurements of known exocyst regulators from membrane fractions isolated from MCF7 cells. (J) Western blot validation for EXOC 1 and 6 from MFC7 total cell lysate, cytosolic fractions, and purified membrane fractions. (K) Western blots examining the levels of Exoc1 and Exoc6 in membrane fractions from MCF7 cells and BT549 cells. (L) Western blots examining Exoc1, Exoc3, and Exoc6 levels in control liver and Myc-O/E hepatic tumors. Student’s t test was used for all pairwise comparisons, and one-way ANOVA was used for all experiments containing >2 sample groups. Error bars represent the standard deviation. Source data are available for this figure: SourceData F4.$

ROS prevents exocyst binding to the membrane. (A) A diagram describing the MCF7 cell model used to examine the impact of a 30-min ROS exposure on the membrane proteome. (B) A volcano plot examining the changes in membrane proteome caused by ROS exposure (FDR < 0.05). (C) A heat map depicting the expression changes of the top 100 most differentially regulated proteins in membranes isolated from ROS-exposed cells (FDR < 0.05). (D) Protein abundances for several common membrane proteins in control and H₂O₂-treated cells. (E) Membrane abundances for exocyst complex components in control and H₂O₂-treated MCF7 cells (FDR < 0.05). (F) A heat map showing the top 100 most differentially regulated proteins in membrane fraction from 3T3 cells and 3T3 treated with H₂O₂. (G) A heat map representing proteomic-based measurements of the detectable subunits of the exocyst complex from membrane fractions isolated from Drosophila eggs. (H) A heat map representing proteomic-based measurements of the detectable subunits of the exocyst complex from membrane fractions isolated from NIH3T3 cells. (I) A heat map representing proteomic-based measurements of known exocyst regulators from membrane fractions isolated from MCF7 cells. (J) Western blot validation for EXOC 1 and 6 from MFC7 total cell lysate, cytosolic fractions, and purified membrane fractions. (K) Western blots examining the levels of Exoc1 and Exoc6 in membrane fractions from MCF7 cells and BT549 cells. (L) Western blots examining Exoc1, Exoc3, and Exoc6 levels in control liver and Myc-O/E hepatic tumors. Student’s t test was used for all pairwise comparisons, and one-way ANOVA was used for all experiments containing >2 sample groups. Error bars represent the standard deviation. Source data are available for this figure: SourceData F4.