Fig 2: Human B7-H4 gene expression is a prognostic factor in HER2+ BCs and limits the therapeutic efficacy of trastuzumab. (A) Quantitation of CD68 and B7-H4 IHC staining in paired samples of HER2+ BC patients before and after neoadjuvant trastuzumab therapy or neoadjuvant chemotherapy without trastuzumab (n = 50 in the trastuzumab group; n = 58 in the chemotherapy group). ***P < 0.001, Student’s t test. (B) Representative images of CD68, B7-H4, CD8, and CD56 in serial sections of the paired clinical samples from HER2+ breast cancer patients before and after neoadjuvant therapy. Scale bar, 50 µm. (C) Kaplan–Meier survival curve for BC patient data and stratified into low and high groups based on the average expression of B7-H4 in BC-infiltrating macrophages. (D) Correlation of infiltrations (number per view field) of B7-H4+ macrophages and NK cells or CD8+ T cells in clinical samples from BC patients after neoadjuvant trastuzumab therapy [n = 50, Pearson’s correlation coefficient (R and P are shown)].

Fig 3: Flow cytometry analysis for CD56 (NCAM1) and CD326 (EPCAM) in undifferentiated RiPS cells as well as ME and MP (p10) cells derived from RiPS cells.MP cells exhibit a similar cell surface expression of these two markers as ME.DOI: http://dx.doi.org/10.7554/eLife.08413.014

Fig 4: Association of CD56 expression in target cells with increased NK-92-induced caspase 3 activation. NK-92 cells were coculture or not with CFSE-labeled target cells for 2 hours at 1:1 effector-to-target ratio then NK-induced and spontaneous active caspase 3 levels in target cells were measured by flow cytometry. (A) Representative histograms showing the percentage of active caspase 3-positive (aCasp3+) target cells. (B) Graph presenting the mean percentages ± SD of NK-induced caspase 3 activation (corrected for spontaneous caspase 3 activation) in target for at least three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 versus hTERT-HME1 target cells.

Fig 5: Flow cytometry gating strategy for quantification of human type II NKT cells. (A) Human T2NKT cells were defined as CD3+ CD56+ CD161+ TCR?d- TCRVa7.2- TCRVa24- lymphocytes. Gating of human PBMC resulted in a very low frequency of circulating T2NKT cells (0.3% of total CD3+ T cells). (B) Gating of human intrahepatic lymphocytes (IHL) revealed a higher frequency of T2NKT cells in non-steatotic liver (6.5% of total CD3+ T cells). Intrahepatic T2NKT cells could be subdivided into CD4+ and CD8+ cells. (C) Frequency of human T2NKT cells with respect to CD3+ T cells from 3 PBMC and 3 liver samples (steatosis 0%, n = 2; steatosis 15%, n = 1).