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. Author manuscript; available in PMC: 2017 Dec 21.
Published in final edited form as: Leukemia. 2016 May 20;30(8):1803–1804. doi: 10.1038/leu.2016.129

Figure 2.

Figure 2

Ibrutinib therapy decreases total protein and mRNA transcript levels of BTK. Primary CLL cells from patients were isolated fresh from peripheral blood samples before treatment (baseline), and at 2, 4 and 12 weeks of 420 mg daily oral dosing of ibrutinib. Blood samples were collected from six patients in green-top tubes and CLL cells were isolated by Ficoll-Hypaque gradient. Cell pellets were used for total BTK protein levels (a) and BTK mRNA levels (b) from six patients with CLL. For the former, cell lysates were prepared and after protein quantitation, immunoblots were run to analyse total BTK protein levels. Immunoblots were quantitated and plotted as a percentage of the control level (baseline sample). For mRNA transcript levels, RT-PCR assay was performed in triplicate from each sample as described previously.8 Student’s t-tests (two-tailed) were performed using the GraphPad Prism6 software (GraphPad Software, Inc., San Diego, CA, USA) to compare values in untreated samples compared with all treated samples and the P-values for the comparisons in a and b are 0.0001 and 0.0003, respectively.