(A) Quantitative RT-PCR analysis of individual let-7 miRNAs in naive CD8 T cells stimulated with plate-bound anti-TCR (μg as indicated) and anti-CD28 (5 μg) for 24 hr, presented relative to results …
Quantification of the expression of miRNAs by qPCR, and flow cytometry analysis of the phenotype of naive CD8 T cells.
(A) Quantitative RT-PCR analysis of individual let-7 miRNAs in naive CD8 T cells stimulated with plate-bound anti-TCRβ (μg as indicated) and anti-CD28 (5 μg) for 24 hr (from Figure 1A), presented …
(A) Quantitative RT-PCR analysis of individual let-7 miRNAs in naive CD8 T cells stimulated with plate-bound anti-TCRβ (10 μg) and anti-CD28 (5 μg) over increasing periods of time as indicated, …
(A) Quantitative RT-PCR analysis of individual let-7 miRNAs in naive and effector (day 5) TCR transgenic CD8 T cells from P14+Rag2−/−, and P14+Lin28TgRag2-/− mice. (B) Surface expression (right) and …
(A) Schematic representation of the hypothesis that let-7 expression inhibits the differentiation of CD8 T cells. (B) Quantification of the number of donor (CD45.2+) CD8 T cells from P14+wild-type …
Quantification of let-7 expression by qPCR, and flow cytometry analysis of effector CD8 T cells during the response to LCMV infection.
(A) Quantitative RT-PCR analysis of individual let-7 miRNAs in naive and effector (day 5) TCR transgenic CD8 T cells from P14+Rag2−/−, and P14+let-7TgRag2−/− mice (B) Analysis of the survival of Rag2…
(A) Analysis of the proliferation of CellTrace Violet-labeled naive CD8 T cells from the indicated mice 72 hr after activation with anti-CD3 mAbs. (B) Quantitative RT-PCR analysis of cell cycle …
Quantification of the expression of cell cycle regulators, Myc and Tfap4, metabolic enzymes, and let-7 miRNAs by qPCR.
(A) Pre- sort and post- sort purities for naive CD8 T cells from wild-type, let-7Tg, and Lin28Tg mice. Cells were isolated from the lymph nodes of the indicated mice, and enriched for T cells using …
(A, B) Cytotoxicity assay of differentiated CTLs from P14+wild type, P14+let-7Tg, and P14+Lin28Tg lymph nodes (A) or from P14+wild-type (WT), or P14+let-7Tg+Lin28Tg+Rag2−/− (4Tg) lymphocytes …
Quantification of cytotoxic assays and expression of effector molecules assessed by qPCR and flow cytometry in CTLs.
Quantitative RT-PCR analysis of effector molecule mRNA expression: Prf1 (Perforin), Gzma (Granzyme A), Gzmb (Granzyme B) in effector CD8 T cells from P14+wild-type, or P14+let-7Tg+Lin28Tg+Rag2−/− …
(A) Quantitative RT-PCR analysis of Eomes (Eomesodermin) and Tbx21 (T-bet) mRNA in naive and effector CD8 T cells (5 days after anti-CD3 mAb stimulation) generated from wild-type, let-7Tg, and …
Quantification of luciferase activity, and the expression of transcription factors in CTLs assessed by flow cytometry and qPCR.
(A, B) Quantitative RT-PCR analysis of Notch target genes: Hes1 (Hes family BHLH transcription factor 1), Hey1 (Hes related family BHLH transcription factor with YRPW motif 1), Heyl (Hes related …
(A) Quantitative RT-PCR analysis of individual let-7 miRNAs in NIH 3T3 fibroblasts, presented relative to results obtained for the small nuclear RNA (U6) (B) Expression analysis of three let-7 …
(A) Quantitative RT-PCR analysis of Eomes mRNA, presented as the fold change in expression, normalized to wild-type (left) and MFI of Eomes protein expression, normalized to wild-type (right), from …
Quantification of cytotoxic assays and expression of effector molecules assessed by qPCR and flow cytometry in CTLs.
Staining of Granzyme B in CTLs generated from the indicated mice. n.s., not significant (p>0.05), 4Tg without Dox compared with 4Tg with Dox using two-tailed Student’s t-test. Data are from one …
(A) Cytotoxicity assay demonstrating specific target lysis of LCMV gp33 or LCMV np396 peptide-pulsed splenocytes by differentiated P14+CD4Cre+Eomesflfl/flLin28Tg CTLs transduced with the indicated …
Upon TCR stimulation, let-7 miRNAs are rapidly down-regulated, in order to release the ‘molecular brake’ let-7 places on CD8 T cell differentiation in naive cells. Our study suggests that let-7 …
A list of all genes containing a 9-base pair let-7 binding motif in either the 3’ untranslanted region (UTR) or open reading frame (ORF) from both the mouse and human genomes, where a (+) or (-) demonstrates the orientation of the gene in the genome.
The ending models are an illustration of the location of the let-7 binding motifs within the human and mouse Eomes gene, where red represents exons and pink represents introns.
Primer sequences and Taqman Assay catalog numbers.