Figure 5. Long-Range Interactions of the Distal V_H Gene Region with the Proximal Igh Domain.

(A) 4C-seq interaction pattern of PAIR8 with sequences across the Igh locus in cultured Pax5^−/−Rag2^−/− and Rag2^−/− pro-B cells.

(B) Long-range interactions of the indicated distal viewpoints (red) with sequences of the Igh 3′ region in the indicated cultured pro-B cell types. See legends of Figures 1 and 2 for further explanations. The data of all viewpoints in (A) and (B) are average values of three experiments except for the PAIR4-HD analysis of Pax5^−/−Rag2^−/− pro-B cells (average of two experiments).

(C) Comparison of the 4C-seq patterns between random BAC libraries and Rag2^−/− pro-B cells (analyzed from J558.89-HD). See legend of Figure 4D for normalization of the BAC sequencing pattern. Yellow shading denotes regions with a 4C-seq pattern that differs between the random BAC libraries and Rag2^−/− pro-B cells. The specificity of the 4C-seq interactions in the IGCR1/D_H region (asterisk) of Rag2^−/− pro-B cells compared to the random BAC library was confirmed by 3C-qPCR (Figure S4E).