Figure 4 miR-155 knockdown in myeloid... | American Society of Hematology

Figure 4

$Figure 4. miR-155 knockdown in myeloid cells reduces TAMs with pro-inflammatory activity. (A) The composition of hematopoietic infiltrate in PyMT breast cancer was determined by flow cytometric analysis of CD45+ cells derived from dissociated tumors. The proportion of Gr1+ cells, CD4+ or CD8+ T lymphocytes, and CD19+ B lymphocytes within the total CD45+ cells is shown (mean ± SEM, n = 9-10). (B) Shown is representative immunofluorescence microscopy of tumor sections (top panels) from 13-week-old PyMT Kd or PyMT Ctrl mice stained for the pan-macrophage marker CD68 and IBA-1, a marker associated with macrophage activation. Original magnification ×20. Shown is the quantification of the CD68+ (bottom, left panel) and IBA-1+ (bottom, right panel) areas from PyMT tumor sections relative to PyMT Ctrl (n = 6 mice from 2 independent experiments). (C) The graph (upper panel) depicts the fraction of CD11c+ TAMs within the total CD45+ hematopoietic tumor infiltrate as measured by flow cytometry. Shown are 2 independent experiments; each point is a pool of 6 and 3 mice in experiment 1 and experiment 2, respectively. The graph lower panel shows the ratio of CD11c/Mrc1 transcript in the CD11b+F4/80+ TAM population as measured by qPCR (n = 3 pools of 3 mice each). Statistical analyses were performed by Student t test. * P < .05.$

miR-155 knockdown in myeloid cells reduces TAMs with pro-inflammatory activity. (A) The composition of hematopoietic infiltrate in PyMT breast cancer was determined by flow cytometric analysis of CD45⁺ cells derived from dissociated tumors. The proportion of Gr1⁺ cells, CD4⁺ or CD8⁺ T lymphocytes, and CD19⁺ B lymphocytes within the total CD45⁺ cells is shown (mean ± SEM, n = 9-10). (B) Shown is representative immunofluorescence microscopy of tumor sections (top panels) from 13-week-old PyMT Kd or PyMT Ctrl mice stained for the pan-macrophage marker CD68 and IBA-1, a marker associated with macrophage activation. Original magnification ×20. Shown is the quantification of the CD68⁺ (bottom, left panel) and IBA-1⁺ (bottom, right panel) areas from PyMT tumor sections relative to PyMT Ctrl (n = 6 mice from 2 independent experiments). (C) The graph (upper panel) depicts the fraction of CD11c⁺ TAMs within the total CD45⁺ hematopoietic tumor infiltrate as measured by flow cytometry. Shown are 2 independent experiments; each point is a pool of 6 and 3 mice in experiment 1 and experiment 2, respectively. The graph lower panel shows the ratio of CD11c/Mrc1 transcript in the CD11b⁺F4/80⁺ TAM population as measured by qPCR (n = 3 pools of 3 mice each). Statistical analyses were performed by Student t test. * P < .05.

This Feature Is Available To Subscribers Only