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. 2020 Oct 23;35(6):1621–1630. doi: 10.1038/s41375-020-01055-7

Fig. 1. ERK1/2 factors in the capacity of Wnt5a to enhance CLL proliferation.

Fig. 1

a Immunoblot analysis of lysates prepared from primary CLL cells (representative of three patients) pretreated without (-) or with (+) cirmtuzumab (20 μg/ml) for 2 h and stimulated without (-) or with (+) Wnt5a (100 ng/ml) for 5 min; expression of total ERK1/2, and activated pERK1/2 was measured, as indicated on the left. b Mean proportions of dividing CLL cells from each of 6 patients (3 U-CLL and 3 M-CLL) under conditions indicated at the bottom. P < 0.05; P < 0.01; P < 0.001, as assessed by 2-tailed Student’s t test. c CLL cells transfected with control siRNA or siRNA targeting ERK1/2 and examined 72 h later by western-blot (representative of 3 patients). Cell viability was ≥90% before siRNA transfection and ≥80% in either control or ERK1/2-siRNA transfected cells. d Mean proportions of dividing CLL cells from each of 6 patients (3 U-CLL and 3 M-CLL) under conditions indicated at the bottom. P < 0.05; P < 0.01; P < 0.001, as assessed by 2-tailed Student’s t test. e Fluorescence of CLL cells stained with CFSE and treated with CD154, without (–) or with (+) Wnt5a. The percentage of dividing cells is indicated in each histogram. f Gene set enrichment analysis (GSEA) on the transcriptomes of paired pre-treatment (Pre-Rx) and post-treatment CLL cells at D28 (D28) evaluating for pre and post-treatment differences in the expression of ERK1/2 downstream target genes. Gene set size (SIZE), normalized enrichment score (NES), and FDR q value (FDR q) are indicated.