Fig. 4.
PCR analysis of the Dβ-Jβ and Vβ-DJβ rearrangements of the TCR gene. (A) Schematic presentations of the PCR-based analyses of β-chain D-J (left) and V-DJ (right) gene rearrangements. (B) and (C) Enriched primitive marrow progenitors were cultured with SF, IL-11, and IL-7 in methylcellulose. On day 11 of culture, resulting primary colonies were individually harvested, pooled, and sorted on the basis of the expression of Ly-6A/E and FcγRII/III. DNA was extracted from crude and sorted cells, control monocyte/macrophage cell line (Raw 264.7) and thymocytes. One hundred micrograms of DNA was PCR amplified for 25 cycles for (Dβ-Jβ, B) or for 35 cycles (Vβ-DβJβ, C). Signals were visualized by using a DIG-conjugated probe and a DIG luminescent detection kit. * Signals were obtained after a short exposure.

PCR analysis of the Dβ-Jβ and Vβ-DJβ rearrangements of the TCR gene. (A) Schematic presentations of the PCR-based analyses of β-chain D-J (left) and V-DJ (right) gene rearrangements. (B) and (C) Enriched primitive marrow progenitors were cultured with SF, IL-11, and IL-7 in methylcellulose. On day 11 of culture, resulting primary colonies were individually harvested, pooled, and sorted on the basis of the expression of Ly-6A/E and FcγRII/III. DNA was extracted from crude and sorted cells, control monocyte/macrophage cell line (Raw 264.7) and thymocytes. One hundred micrograms of DNA was PCR amplified for 25 cycles for (Dβ-Jβ, B) or for 35 cycles (Vβ-DβJβ, C). Signals were visualized by using a DIG-conjugated probe and a DIG luminescent detection kit. * Signals were obtained after a short exposure.

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