Dataset on the changes of neutrophils treated with retinoic acid

The data presented in this article are related to the research article entitled “Retinoic acid induces hypersegmentation and enhances cytotoxicity of neutrophils against cancer cells” (S. Shrestha, S.Y. Kim, Y.J. Young, J.K. Kim, J.M. Lee, M. Shin, D.K. Song, C.W. Hong, 2017) [1]. This article complements the potential of retinoic acid to induce changes in effector function of human neutrophils. Here the datasets describe the rate of apoptosis, changes in numbers of nuclear lobes, and the expressions of surface markers in human neutrophils in presence or absence of retinoic acid. The tumor growth in recipient mice with adoptive transfer of retinoic acid-treated neutrophils was evaluated. The included data is made publicly available to criticism and extended analysis.

. This article complements the potential of retinoic acid to induce changes in effector function of human neutrophils. Here the datasets describe the rate of apoptosis, changes in numbers of nuclear lobes, and the expressions of surface markers in human neutrophils in presence or absence of retinoic acid. The tumor growth in recipient mice with adoptive transfer of retinoic acidtreated neutrophils was evaluated. The included data is made publicly available to criticism and extended analysis.
& Neutrophils were exposed to retinoic acid

Experimental features
Apoptosis, nuclear segmentation, and surface marker expressions of neutrophils were determined. Tumor growth in recipient mice was examined.

Data source location
Kyungpook National University, Daegu, Republic of Korea,

Data accessibility
Data are presented in this article

Value of the data
The data highlights the changes in the function of human neutrophils after treatment with retinoic acid.
The data can be useful to compare the rate of apoptosis in neutrophils induced by retinoic acid. The data shows changes in the nuclear morphology and surface expression markers induced by retinoic acid treatment and the effect of adoptive transfer of retinoic-acid-treated donor neutrophils on tumor growth in recipient mouse.
The data can be useful for other researchers investigating the effects of retinoic acid on neutrophils.

Neutrophil isolation
Human blood experiments were approved by the Institutional Research Board of Hallym University and Kyungpook National University. Neutrophils were isolated from venous blood by double density gradient method using histopaque 1077 and 5% dextran [2].

Apoptosis detection
Neutrophils (1 Â 10 6 cells/ml) were treated with retinoic acid (100 nM) for the indicated time. Neutrophils were collected and stained with annexin V/propidium iodide following instructions of apoptosis detection kit. Then cell apoptosis was detected using flow cytometer.

Wright-Giemsa staining for mean lobe count
Hypersegmentation in neutrophils was determined by Wright-Giemsa staining. Isolated neutrophils (2 Â 10 6 ) were treated with retinoic acid (100 nM, Sigma-Aldrich) for 4 h. Cells were then cytospun and stained with hemacolor stain (Merck Millipore) and mean lobe counts were determined.

Flow cytometry
Neutrophils were fixed and then stained. Antibodies targeted against different surface receptors: CD15, CD14, CD16 and CD64 and adhesion molecules: CD177, CD62L, and OLMF-4: FITC conjugate were used. The cells were washed and dissolved in staining buffer. Data acquisition was done using flow cytometer (BD FACS Calibur) and data analyzed by Flowjo.

Adoptive transfer of retinoic acid-treated neutrophils
Neutrophils from naïve mice were harvested by positive selection using Ly6G þ MACS kit. The neutrophils were treated with retinoic acid (100 nM) for 4 h. Recipient BALB/c mice (five weeks old, female) were injected with 1 Â 10 5 4T1 cells in the right leg. Then, recipient tumor-bearing mice were administered retinoic acid treated neutrophils (5 Â 10 6 cells) on day 13 and 16 post tumor injection. Tumor growth was measured every three days until day 21. Fig. 4. Tumor growth in recipient mice with adoptive transfer of retinoic acid-treated neutrophils. Neutrophils were isolated from bone marrow of naïve mice and further treated with retinoic acid (100 nM). Neutrophils were intratumorally injected to recipient tumor bearing mice on day 13 and 15. The relative percentages of tumor growth compared to vehicle-injected mice were calculated. All results are shown as mean 7 SEM (n¼ 4 mice for each group).

Statistical analysis
All statistical analyses were performed using GraphPad prism 5.0 (Graphpad software). AVOVA analysis or two-tailed Student's t-test were performed to determine the significance.