CC-115 reverts CD40-induced chemoresistance. (A-I) CLL cells were cultured on 3T3 fibroblasts or CD40L-expressing fibroblasts (CD40L) in the presence or absence of 1 µM CC-115, CC-214, CC-292, or idelalisib for 3 days. (A) Protein lysates were probed for pS6 (S240/244), peIF4E (S209), p4EBP1 (T37/46), and actin as loading control. Blots from 2 representative CLL samples are shown (patients 70 and 37). (B) Survival was analyzed by DiOC6 staining. Results are shown as mean ± SEM. *P < .05, **P < .01 (1-way ANOVA; n = 10; patients 6, 25, 30, 33B, 34A, 43B, 72-74, and 75). (C) After 3 days, fludarabine sensitivity assay was performed. Apoptosis was assessed by DiOC6/PI staining, and specific apoptosis is shown as mean ± SEM (n = 9; patients 6, 25, 30, 33B, 34A, 43B, and 72-74). (D) After 3 days, an ABT-199 sensitivity assay was performed. Results are shown as mean ± SEM (n = 3; patients 33B, 35B, and 45). (E) Protein lysates were probed for Bim and actin as loading control. Blots of 2 representative CLL samples are shown of 7 analyzed. (F) Protein lysates were probed for Mcl-1, Bcl-XL, Bfl-1, and actin as loading control. Blot from 1 representative CLL sample is shown of 9 analyzed. (G-I) Densitometric analysis of (G) Mcl-1/actin, (H) Bcl-XL/actin, and (I) Bfl-1/actin level is shown. Bars represent mean ± SEM. *P < .05, ** P < .01 (1-way ANOVA).
