چکیده (انگلیسی):

Human basophils require MTs to induce IL-4 gene transcription. (AeC) Levels of IL4, MT1F, MT1X, and MT2A transcripts in blood basophils were measured relative to 18S rRNA
using real-time RT-PCR. Correlations between MT1F and IL4 (A), MT1X and IL4 (B), and MT2A and IL4 (C) expression are shown. Basophil purity was >95%. Student's t-tests were used
for statistical analyses and P-values are indicated. R-values were determined using Pearson's correlation tests. (DeE) Human basophils isolated from 200 ml of venous blood
collected from each healthy donor were transfected with siRNA (SigmaeAldrich) using a two-step electroporator. The viability of the transfected cells ranged from 41% to 46%.
(D) Real-time RT-PCR analysis of MT1F, MT1X, and MT2A transcripts in MT-knock down and control basophils relative to 18S rRNA. (E) MT-knock down (closed bars) and control
(open bars) basophils were stimulated with anti-human IgE for 2 h. IL4 transcript levels (normalised to 18S rRNA) were measured using real-time RT-PCR. Data represent
means ± SD; **P < 0.01 by Student's t-tests compared with control basophils. <D.L., below detection limit. Data representative of two experiments are shown. The primers
were used for PCR reactions as listed in Supplementary Table 1. The primers, except for18S rRNA, were designed and synthesized by TAKARA.