All mouse studies have been conducted according to Animal Welfare Act and the Public Health Service Policy and approved by Vanderbilt University Institution Animal Care and Use Committee (IACUC) and NCI. The animals were housed in pathogen-free units, in compliance with IACUC regulations. C57BL/6 J and LysMCre mice were purchased from Jackson Labs. Mice with a floxed Cebpa gene, called Cebpaflox/flox, were developed as described26 (link). We generated mice with myeloid-specific deletion of Cebpa by breeding Cebpaflox/flox mice to LysMCre mice, which express Cre recombinase under the control of murine lysozyme M promoter. C/EBPαflox/flox;LysMCre (+/−) (CebpaΔ/Δ) and littermate control mice C/EBPαflox/flox; LysMCre (−/−) (WT) were used in the study.
The 32D myeloid cell line, Lewis lung cancer derivative cell line (3LL), B16 melanoma cell line and HUVECs were purchased from ATCC and maintained per standard cell culture techniques.
B16 or 3LL cells (5 × 105 cells), with or without purified Gr1+CD11b+ cells (0.5 × 105 cells), were injected subcutaneously (s.c.) into the left flank of C57Bl/6 mice. The size of tumors was determined by measurement of tumor dimensions at 2–3 day intervals using a caliper. The equation volume = length × (width)2 × 0.5 was used to calculate tumor volume.
The 32D myeloid cell line, Lewis lung cancer derivative cell line (3LL), B16 melanoma cell line and HUVECs were purchased from ATCC and maintained per standard cell culture techniques.
B16 or 3LL cells (5 × 105 cells), with or without purified Gr1+CD11b+ cells (0.5 × 105 cells), were injected subcutaneously (s.c.) into the left flank of C57Bl/6 mice. The size of tumors was determined by measurement of tumor dimensions at 2–3 day intervals using a caliper. The equation volume = length × (width)2 × 0.5 was used to calculate tumor volume.