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(d) Western blot analysis for levels of ETV4 and ETV5 in sorted splenic CD4+PD-1+CXCR5+ TFH cells.
(a) qRT-PCR analysis for levels of Pea3 group genes in splenic CD4+ T cells from Cicf/f (WT) and Cicf/fCd4-Cre (cKO) mice.
(e) Western blot analysis for levels of CIC and ETV5 in sorted non-TFH and TFH cells from spleen of WT C57BL/6 (B6) mice.
(c) qRT-PCR analysis for levels of Etv4 and Etv5 in sorted CD4+PD-1−CXCR5− non-TFH and CD4+PD-1+CXCR5+ TFH cells.
(a) qRT-PCR analysis for levels of TFH-related genes in sorted non-TFH and TFH cells from Cicf/f and Cicf/fCd4-Cre mice.
The analysis was conducted by two of the researchers independently, with analysis for levels of agreement.
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One is a macro-level analysis that measures how domain shift affects corpus-level evaluation; the second is a micro-level analysis for word-level errors.
(e) Western blot analysis for MAF levels in CD4+ T cells infected with control or ETV5-expressing retrovirus.
(d) qRT-PCR analysis for Maf levels in CD4+ T cells infected with control or ETV5-expressing retrovirus in the absence of IL-6 and IL-21.
Western blot analysis for CIC levels in CD4+ T cells during T-cell activation by α-CD3/CD28 antibodies (a) and in sorted naïve (CD44loCD62Lhi) and effector/memory (CD44hiCD62Llo) CD4+ T cells (b).
qRT-PCR analysis for mRNA levels of Pea3 group genes (Etv1, Etv4 and Etv5) (Fig. 5a) and quantitative PCR (qPCR) analysis for Pea3 group gene promoter regions in total CIC-associated DNA fragments (Fig. 5b) verified that Etv4 and Etv5 are direct targets of CIC in CD4+ T cells.
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