Fig 1: Deletion of Aspartyl and Glutamyl Aminopetidases DNPEP and ENPEP inhibits sequential removal of acidic amino acid residues from beta actin N-terminusQuantification of N-terminal processing of beta actin from WT, DNPEP-KO, ENPEP-KO and double knockout HAP1 cells, presented as fraction of the total (left) and as average intensity ratio across samples (right) similarly to that shown in Figure 3. Error bars in the left chart represent SEM, n = 3 independently derived cell lines for each genotype; see Table S4 for the actual values. Individual intensity ratios in the right chart are normalized to WT and plotted for each repeat. See Table S2 for the raw intensity values.
Fig 2: ENPEP knockout results in reduced cell spreading in HAP1 cellsLeft, violin plots of the area occupied by spread cells on the substrate; right, violin plots of cell volume, derived from the measurements of trypsinized spherical cells in suspension. Error bars represent SEM, n = 31 for WT, 30 for ENPEPKO, 29 for DNPEP KO, and 32 for the double knockout for the cell area measurement, and 149 for WT, 156 for ENPEPKO, 166 for DNPEP KO, and 139 for the double knockout for the cell volume measurement. p values were calculated by unpaired 2-tailed Student’s t test, ns: p > 0.05.
Fig 3: DNPEP and ENPEP knockout results in changes in actin polymer levels in HAP1 cellsTop, violin plots of F-actin per unit area (left) and F-actin per cell (right) detected by Phalloidin-AlexaFluor488 staining of different genotypes; bottom, representative images of different genotypes stained with Phalloidin-AlexaFluor488 (scale bar, 10 μm). n = 31 for WT, 30 for ENPEPKO, 28 for DNPEP KO, and 31 for the double knockout for the intensity per cell measurement, and 31 for WT, 30 for ENPEPKO, 29 for DNPEP KO, and 32 for the double knockout for the intensity per unit area measurement. Data is presented as violin plots with median and quartiles. p values were calculated by unpaired 2-tailed Student’s t test, ns: p > 0.05.
Fig 4: DNPEP and ENPEP knockout results in changes in the number and length of filopodia in HAP1 cellsTop, Representative images of cells from each genotype stained with Phalloidin-AlexaFluor488 (white) and DAPI (blue) (scale bar, 10 μm); bottom, violin plots of filopodia number (left) and length (right). n = 181 for WT, 259 for ENPEPKO, 222 for DNPEP KO, and 205 for the double knockout for the filopodia length measurement, and 84 for WT, 84 for ENPEPKO, 71 for DNPEP KO, and 65 for the double knockout for the filopodia numbers measurement. p values were calculated by unpaired 2-tailed Student’s t test, ns: p > 0.05.
Supplier Page from R&D Systems, a Bio-Techne Brand for Aminopeptidase A/ENPEP Protein
Available conjugates: Sizes Available: 10 ug