Analysis of SAP expression in dNK cells. (A) Analysis of SAP mRNA transcript in dNK and pNK cell clones. Total RNA, extracted from dNK and pNK cell clones and converted in cDNA, was subjected to TaqMan-PCR analysis to assay the quantity of SAP mRNA. All values have been normalized to GAPDH transcript and the quantitative PCR (Q-PCR) data are represented as time-fold the normalized SAP transcript detected in the NK92 cell line (used as reference cell line and therefore indicated as 1). Each spot is the mean of 2 independent experiments of Q-PCR analysis performed in triplicates. Statistical analysis of data sets was annotated as follows: *P = .01; **P < .01; and n.s., not significant. (B) Postnuclear cell lysates derived from 1 × 106 dNK, pNK, and XLP-NK cells were spotted on nitrocellulose using a slot-blot system. Membranes were probed with rabbit antisera specific for either actin or SAP. Membranes were then scanned and densitometric analysis of the bands was performed. Densities of actin and SAP in dNK and XLP-NK cells are expressed as percentage of control pNK cells. Data are representative of 3 independent experiments. (C) FACS analysis of SAP expression. Permeabilized rIL-2–cultured XLP-NK, dNK, and pNK cells were stained with the SAP-specific rabbit antiserum. Gray profiles represent negative controls (ie, the antiserum background detected in SAP-deficient XLP-NK cells).
