Cell Culture and Treatment Protocols

The SK-N-SH (Non-MYCN amplification) and SK-N-BE(2) cell line (MYCN amplification) (obtained from the Shanghai Institute of Biological Cell Research, Chinese Academy of Sciences) was cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS; Gibco) [22 (link), 23 ]. Additionally, the THP-1 cell line (also obtained from the Shanghai Institute of Biological Cell Research, Chinese Academy of Sciences) was cultured in 1640 medium supplemented with 10% fetal bovine serum (FBS; Gibco) and 100 U/ml penicillin–streptomycin. HUEVC Human vascular endothelial cells were isolated by collagenase digestion of the human umbilical cord veins obtained from Children's Hospital Affiliated with Chongqing Medical University with the informed consents signed by donors. Human vascular endothelial cells were cultured in Endothelial Cell Medium (Lonza). 0.99 μm BKM 120 for SK-N-SH, 1.2 μm BKM 120 or SK-N-BE(2) (one of a PI3K inhibitor) (MCE)was used for the PI3K-AKT experiment [24 (link)]. For THP-1 cells, 100 ng/ml of PMA (Phorbol 12-myristate 13-acetate) (Multi-Science) was added, and recombinant human CXCL14 (MCE) was used for the experiment in vitro [25 (link)].

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Wu X., Mi T., Jin L., Ren C., Wang J., Zhang Z., Liu J., Wang Z., Guo P, & He D. (2024). Dual roles of HK3 in regulating the network between tumor cells and tumor-associated macrophages in neuroblastoma. Cancer Immunology, Immunotherapy : CII, 73(7), 122.

Publication 2024

FBS Endothelial Cell Medium PMA Phorbol 12-myristate 13-acetate

Corresponding Organization :

Other organizations : Children's Hospital of Chongqing Medical University, Chongqing Medical University

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Variable analysis

independent variables

BKM 120 (PI3K inhibitor) (0.99 μm for SK-N-SH, 1.2 μm for SK-N-BE(2))
PMA (Phorbol 12-myristate 13-acetate) (100 ng/ml) for THP-1 cells
Recombinant human CXCL14 for THP-1 cells

dependent variables

PI3K-AKT pathway in SK-N-SH and SK-N-BE(2) cell lines
Unspecified outcomes for THP-1 cells with PMA and CXCL14

control variables

DMEM medium supplemented with 10% FBS for SK-N-SH and SK-N-BE(2) cell lines
RPMI-1640 medium supplemented with 10% FBS and 100 U/ml penicillin–streptomycin for THP-1 cell line
Endothelial Cell Medium (Lonza) for HUEVC Human vascular endothelial cells

controls

Positive control: None specified
Negative control: None specified

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