PKRA7 blocks endothelial cell branching, myeloid cell migration, and PK2-induced expression of specific chemokine/chemokine receptors.

<p>(<b>A</b>) IHMVECs were plated on Matrigel in indicated treatment groups. Representative photographs were taken at 8 hours after plating. (<b>B</b>) Average number of connections between cells was counted and analyzed. Results are normalized data from 3 independent experiments. Addition of PKRA7 significantly blocks PK2-induced capillary branching (*p≤0.05). (<b>C</b>) 1×10⁵ THP-1 cells on top chamber of transwells were allowed to migrate for 4 hours. Cells were fixed, stained and the number of cells per field of view counted. Results are the normalized average of 3 independent experiments. Addition of PKRA7 significantly blocked PK2-induced monocyte migration (*p≤0.05). (<b>D</b>) 7.5×10⁴ RAW264.7 cells on top chamber of transwells were allowed to migrate for 18 hours. Cells were fixed, stained and the number of cells per field of view counted. Results are the normalized average of 3 independent experiments. Addition of PKRA7 significantly blocked PK2-induced macrophage migration (*p≤0.05). (<b>E</b>) Average measured luminescence of tumor site after IP injection of luciferase-labeled RAW cells into control (n = 4) or PKRA7 (n = 4) treated mice with SC AsPc-1 tumors 30 days after implantation. Average total luciferase counts were lower in mice treated with PKRA7 compared to control (*p≤0.05). (<b>F</b>) qPCR assay to measure the effect of PKRA7 on the expression of chemokines and chemokine receptors that were identified to be induced by PK2 treatment. Data on the mRNA level changes were shown as log2 of the Ct value changes. PKRA7 inhibits upregulation of CCL27, CCR10, CCR4, CCR5, and CCR6 (*p≤0.05).