Figure S5. ROS production disrupts... | Rockefeller University Press

Figure S5.

$ROS production disrupts endosomal traffikcing and cell cell communication. (A) GO enrichment for protein domains found in proteins downregulated in membrane fractions purified from MCF7 cells exposed to ROS. (B) ROS levels from MCF7 cells and BT549 cells. (C) Quantification of the levels of EXOC1 and EXOC6 in membrane fractions from MCF7 cells and BT549 cells (n = 3). (D) A heat map showing the abundance of RAB family GTPases in purified membrane fractions from MCF7 cells. (E) DHE staining of PHB1-RNAi and PHB1-RNAI; +RAB11-O/E ovarioles. (F) quantification of peb antibody staining fluorescence from control and PIPK-RNAi ovarioles (n = 20). (G) volcano plot depicting the protein level differences between control 3T3cell and 3T3 cells cultured with H2O2. (H) A heat map depicting the expression of NRF2 target genes in cells adjacent to ROS-producing cells. (I) a heatmap of the gene expression abundance changes in genes associated with the term “secreted protein.” Expression levels are expressed as a ratio ROS-coculture/control. (J) A heatmap of the top 100 most significant differentially regulated genes in cells adjacent to ROS-producing cells. (K) A heatmap of the abundance of genes associated with the gene ontology term “extracellular protein” from our list of genes downregulated in cells adjacent to ROS-producing cells(Red text indicates a known role in cancer growth, metastasis, and angiogenesis). (L) ROS-Levels in Hek293 control cells, DAAO-expressing, and cells co-cultured adjacent to ROS-producing DAAO+ cells (n = 25). (M) A Venn diagram showing the overlap of genes known to be regulated by extracellular ROS exposure (NCBI GEO: GSE227554, H2O2 exposure for 6 h) and our list of genes regulated by co-culture in contact with ROS-producing cells. (N) GO term enrichment for the 23 genes overlapping our data and the H2O2-regulated genes studied in GSE227554. 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.$

ROS production disrupts endosomal traffikcing and cell cell communication. (A) GO enrichment for protein domains found in proteins downregulated in membrane fractions purified from MCF7 cells exposed to ROS. (B) ROS levels from MCF7 cells and BT549 cells. (C) Quantification of the levels of EXOC1 and EXOC6 in membrane fractions from MCF7 cells and BT549 cells (n = 3). (D) A heat map showing the abundance of RAB family GTPases in purified membrane fractions from MCF7 cells. (E) DHE staining of PHB1-RNAi and PHB1-RNAI; +RAB11-O/E ovarioles. (F) quantification of peb antibody staining fluorescence from control and PIPK-RNAi ovarioles (n = 20). (G) volcano plot depicting the protein level differences between control 3T3cell and 3T3 cells cultured with H₂O₂. (H) A heat map depicting the expression of NRF2 target genes in cells adjacent to ROS-producing cells. (I) a heatmap of the gene expression abundance changes in genes associated with the term “secreted protein.” Expression levels are expressed as a ratio ROS-coculture/control. (J) A heatmap of the top 100 most significant differentially regulated genes in cells adjacent to ROS-producing cells. (K) A heatmap of the abundance of genes associated with the gene ontology term “extracellular protein” from our list of genes downregulated in cells adjacent to ROS-producing cells(Red text indicates a known role in cancer growth, metastasis, and angiogenesis). (L) ROS-Levels in Hek293 control cells, DAAO-expressing, and cells co-cultured adjacent to ROS-producing DAAO+ cells (n = 25). (M) A Venn diagram showing the overlap of genes known to be regulated by extracellular ROS exposure (NCBI GEO: GSE227554, H₂O₂ exposure for 6 h) and our list of genes regulated by co-culture in contact with ROS-producing cells. (N) GO term enrichment for the 23 genes overlapping our data and the H₂O₂-regulated genes studied in GSE227554. 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.