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. 2023 Jan;29(1):30–43. doi: 10.1261/rna.079472.122

FIGURE 2.

FIGURE 2.

A-Myb expression precedes that of Tcfl5. (A,B) Temporal A-Myb and Tcfl5 mRNA expression in a seminiferous tubule section from adult mouse testis was detected. Two-color RNA fluorescent in situ hybridization (RNA-FISH) (A). Single-color RNA-FISH (B). (C) Strategy to generate Tcfl5 knockout mice using a single guide RNA (sgRNA) and Cas9. Scissors indicate sites targeted by sgRNAs designed to delete Tcfl5 exons 2 and 3. RNA-seq was used to measure the abundance of Tcfl5 mRNA in adult testes. (D) Abundance of TCFL5 protein in Tcfl5em1/em1 homozygous and Tcfl5+/em1 heterozygous mutant testes, relative to C57BL/6 wild-type testes. ACTIN serves as a loading control. Each lane contained 50 µg testis protein. (E) Abundance of A-MYB and TCFL5 proteins in A-Myb−/− and Tcfl5em1/em1 homozygous mutant mouse testes. ACTIN serves as a loading control. Each lane contained 50 µg testis protein. (F) A-MYB and TCFL5 ChIP-seq peaks at the promoters of the A-Myb and Tcfl5 genes.