. Author manuscript; available in PMC: 2019 Mar 1.

Published in final edited form as: J Immunol Methods. 2017 Dec 16;454:27–31. doi: 10.1016/j.jim.2017.12.001

Table 2. Quantitative RT-PCR amplification of cDNA prepared from RNA isolated from live CD11c^-Gr1^-/loMHCII^- eosinophils.

The housekeeping gene, beta-glucuronidase (gusB) was amplified from first-strand cDNA (+RT) generated from RNA samples shown in Fig. 3B prepared from FACS-isolated eosinophils from lungs of IL5tg mice and mice sensitized and challenged with A. fumigatus (Af) shown in Fig. 3B. No amplification product was detected from RNA samples in which reverse transcriptase was omitted (-RT) or in the absence of RNA template (ntc, no template control); undet, undetected, C_t > 40 cycles.

Well	Detector	cDNA sample	C_t (cycles)
B1	gusb	IL5tg +RT	27.00
B2	gusb	IL5tg +RT	26.98
B5	gusb	IL5tg - RT	undet
B6	gusb	IL5tg - RT	undet
B10	gusb	ntc	undet
D1	gusb	Af + RT	27.09
D2	gusb	Af + RT	27.10
D5	gusb	Af - RT	undet
D6	gusb	Af - RT	undet
D10	gusb	ntc	undet

Table 2. Quantitative RT-PCR amplification of cDNA prepared from RNA isolated from live CD11c-Gr1-/loMHCII- eosinophils.

Table 2. Quantitative RT-PCR amplification of cDNA prepared from RNA isolated from live CD11c^-Gr1^-/loMHCII^- eosinophils.