Anti cxcr4

FACS with anti-c-Kit (Invitrogen; cat. no. CD11705), anti-CXCR4 (BioLegend; clone 12G5, cat. no. 306506), and anti-EpCAM (Invitrogen; clone G8.8, cat. no. 17-5791-80) was used to detect DE cells. Basically, cells were incubated with antibodies for 30 min at 4°C, followed by being washed and suspended in 0.1% BSA/PBS buffer. PE and APC filters were then used to detect cells double positive for Kit and CXCR4 or EpCAM and CXCR4 by signal intensity gating. FACS with anti-CGRP antibody (Abcam; clone 4902, cat. no. ab81887) was used to detect CGRP⁺ cells. Cells were first incubated with anti-human CGRP antibody for 30 min at room temperature followed with incubation of secondary antibody conjugated with R-PE for 30 min at room temperature. Then cells were washed and suspended in 0.1% BSA/PBS buffer. PE filter was then used to separate cells into CGRP⁺ and CGRP⁻ subgroups by signal intensity gating. Negative controls stained with control IgG instead of primary antibodies were always performed with sample measurements. Flow cytometry machine of BD FACSAria II and software of Flowjo were mainly used to collect and analyze the flow cytometry data.