Figure 2. Identification of cells with... | Rockefeller University Press

Figure 2.

$Figure 2. Identification of cells with BRAF-V600E mutation in LCH lesions, circulating cells, and BM aspirates. (A) gDNA was isolated from biopsies of LCH lesions, and the percentage of cells with the BRAF-V600E allele was estimated by qPCR (line = median, 8.0%). (B) gDNA was isolated from PBMCs of patients with active LCH before initial therapy or at a time of recurrence before salvage chemotherapy. The percentage of circulating cells with the BRAF-V600E allele in patients from different clinical risk categories was estimated by qPCR. (C) Estimate of recurrence by circulating BRAF status among those who were clinically classified with low-risk LCH (blue = no circulating cells with BRAF-V600E detected, red = circulating cells with BRAF-V600E detected). (D) gDNA was isolated from gradient-separated leukocytes from synchronous peripheral blood and BM aspirate collected from patients with active LCH. The percentage of cells with the BRAF-V600E allele was determined by qPCR. (Every bar represents a single blood sample; technical duplicates were used in all experiments.) (E) The percentage of circulating cells with the BRAF-V600E allele was determined by qPCR of serial PBMC samples in one patient with disease refractory to multiple salvage therapies (red), and finally cured with clofarabine (green). (Every data point represents a single blood sample; technical duplicates were used in all experiments.)$

Identification of cells with BRAF-V600E mutation in LCH lesions, circulating cells, and BM aspirates. (A) gDNA was isolated from biopsies of LCH lesions, and the percentage of cells with the BRAF-V600E allele was estimated by qPCR (line = median, 8.0%). (B) gDNA was isolated from PBMCs of patients with active LCH before initial therapy or at a time of recurrence before salvage chemotherapy. The percentage of circulating cells with the BRAF-V600E allele in patients from different clinical risk categories was estimated by qPCR. (C) Estimate of recurrence by circulating BRAF status among those who were clinically classified with low-risk LCH (blue = no circulating cells with BRAF-V600E detected, red = circulating cells with BRAF-V600E detected). (D) gDNA was isolated from gradient-separated leukocytes from synchronous peripheral blood and BM aspirate collected from patients with active LCH. The percentage of cells with the BRAF-V600E allele was determined by qPCR. (Every bar represents a single blood sample; technical duplicates were used in all experiments.) (E) The percentage of circulating cells with the BRAF-V600E allele was determined by qPCR of serial PBMC samples in one patient with disease refractory to multiple salvage therapies (red), and finally cured with clofarabine (green). (Every data point represents a single blood sample; technical duplicates were used in all experiments.)