Fig. 2: Characterisation of ULK1 inhibition in GCB cell lines and provides a rationale for its targeting in GCB-DLBCL patients.

A Gene Ontology (GO) analysis of downregulated genes in Oci-Ly1 (MRT68921-treated with 2.5 µM vs. vehicle). B GSEA representing negative NES values of genes downregulated associated to regulation of gene expression, c-MYC transcriptional activation and c-MYC pathway. C GCB Oci-Ly1 and Oci-Ly18 cell lines treated with MRT68921 for 0.5, 1 and 2 h and phosphorylation of c-MYC Ser62 was measured. This blot represents three independent experiments. The relative protein expression was normalised to the total protein and a fold was generated. One-way ANOVA with post hoc Dunnett’s multiple comparisons test. Representative error bars of mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. D Endoplasmic reticulum stress is induced in response to autophagy inhibition. GCB Oci-Ly1 and Oci-Ly18 cells were treated with 2.5 µM MRT68921 in a time-dependent manner. Activation of the PERK pathway was confirmed by phosphorylation of eIF2a and activating transcription factor ATF-4. Immunoblotting was used for detection. E GSEA compared autophagy transcripts between gene expression profiling (GEP) of GCB (n = 240) and ABC (n = 121) patients assigned to R-CHOP treatment arm. Venn diagram illustrating the co-expression of differentially expressed genes from the ULK1 and VPS34 complexes in GCB patients. Increased gene expression of the ULK1 complex is associated with poor overall survival upon R-CHOP treatment. Transformed p values from (Supplementary Fig. 9A) were plotted, data presents mean ± SD and student t test unpaired two tailed, *P < 0.05.