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. 2018 Sep 14;5(6):285–300.

FIG. 3.

FIG. 3

Repression of TCRβ chain mRNA in T × L-cell hybrids. (A) Total RNA (15 μg) from parental and hybrid cells were electrophoresed through formaldehyde/agarose gel, transferred to Nytran membranes, and hybridized with a labeled TCR-Cβ radioactive probe. Lanes 1–10: BW5147, L, Hy1A, Hy2A, Hy3A, Hy1B, Hy3B, Hy4B, Hy1C, and Hy3C, repectively. (B) Total BW5147 RNA (10, 5, 2.5, and 1 μg; lanes 1–4, respectively), poly(A)+ mRNA of Hy1A (2, 5, and 10 μg; lanes 5–7, respectively), and Hy3C poly(A)+ mRNA (1, 2, and 3 μg; lanes 8–10, respectively) were hybridized with a labeled TCR-Cβ chain probe. The blots were stripped and rehybridized with a β-actin cDNA probe. The blots containing the β-actin transcripts are shown in the lower parts of this figure. (C) Analysis of germ line and rearranged TCRβ chain genes in T × L cell hybrids. DNA of parental [L and T (BW5147)] and hybrid cells (15 μg) was digested with EcoRI, electrophoresed on 1% agarose gel, and transfered to Nytran paper. The blot was hybridized to a TCR-Cβ probe and autoradiographed. Hybrids 1–5 are Hy2A, Hy3A, Hy1B, Hy3B, and Hy2C respectively. Bands 4.4 kb and 2.3 kb represent the germ line TCR-Cβ gene and the 2.0-kb band represent the productively rearranged TCR-Cβ in T(BW5147) cells. Note that all the hybrids contain the 2.0-kb band.