Fig. 2: Lymphoma cell lacking 4E-BP1 are resistant to TOR-KI yet are sensitive to SBI-756 treatment.

OCI-LY1 cells stably expressing Cas9 were transfected with guide RNAs. Clones were isolated that carried an empty vector (a, b, e); or sgRNAs specific for 4E-BP1 (4E-BP1 KO) (c, d, f). Parental OCI-LY1 cells gave comparable results to EV transfected OCI-LY1 (data not shown). a–d. Cells were treated for 48 h with titrated amounts of venetoclax without (vehicle) or with MLN0128 (a, c) or SBI-756 (b, d). Viability was assessed using annexin V and PI staining. n = 4. *p < 0.05, ***p < 0.01, ****p < 0.005, ns not significant. Statistics were done using Two-way ANOVA. e, f PLA comparing parental OCI-LY1 to 4E-BP1 KO cells after 4 hours treatment. Five fields were imaged and quantified by a blinded observer (e). Data are plotted for individual experiments with the means of each group indicated by a horizontal line (f). n = 3. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001, ns not significant. Paired one-sample t-test vs. control. g Viability assay of VAL cells (naturally lacking 4E-BP1⁶) following 48 h of treatment with venetoclax, MLN0128 or SBI-756 (all within 100–1000 nM range). h PLA comparing OCI-LY1 to VAL cells (naturally lack 4E-BP1⁶) following 4 h of treatment. Fields imaged and quantified by a blinded observer. Data are plotted for individual experiments with the means of each group indicated by a horizontal line. n = 3. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001, ns not significant. Paired one-sample t-test vs. control.