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Microfluidic qRT-PCR (Applied Biosystems): included 93 apoptotic genes; 384 well plate

Detailed protocol for handling CLL lymphocytes, including gene expression analysis of apoptotic genes using microfluidic qRT-PCR and 384-well plate Real-time RT-PCR. Methods for assessing Bcl-2 family members and combination therapy efficacy discussed.

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Microfluidic qRT-PCR (Applied Biosystems): included 93 apoptotic genes; 384 well plate

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  1. Supplemental Table 1. Details of handling and experimental usage of CLL lymphocytes obtained from patients Microfluidic qRT-PCR (Applied Biosystems): included 93 apoptotic genes; 384 well plate Real-time RT-PCR : Real-time reverse-phase polymerase chain reaction RPPA: Reverse Phase Protein Array

  2. Supplemental Table 2. Cytogenetics and prognostic markers of patient samples used for RPPA, immunoblot analyses. NA: Information not available; PR: Partial response; SD: Stable disease; PROG: Progressive disease; M: Mutated; UM: Unmutated

  3. Supplemental Table 3. List of ligands, reagents and antibodies along with their sources

  4. Supplemental Table 4. Transcript and protein expressions of Bcl-2 family members measured through different techniques. *Unpaired student t-test; ND, below limit of detection; NA, not applicable (not in the assay)

  5. Supplemental Table 5. Combination of duvelisib and venetoclax: Expected % apoptosis vs Observed % apoptosis . Red: Expected % apoptosis < Observed % apoptosis indicates either additive or synergistic combination. Green: Expected % apoptosis > Observed % apoptosis indicates either non-additive, non-synergistic or antagonistic combination. Method of calculating Expected % apoptosis: 1) Raw viability was normalized to "% viability”. 2) Expected % viability was calculated using following formula: “ % viability of drug A * % viability of drug B / 100” . 3) Expected % apoptosis was calculated as "100-(Expected % apoptosis)"

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