Fig 2: Upregulation of AATs in CRC patient samples and their association with oncogenic KRAS mutations: (A–C) Box‐whisker plots show mRNA expression levels of AATs (SLC1A5, SLC7A5, and SLC38A2) in TCGA colon adenocarcinoma patient samples (n = 286) and normal samples (n = 41). Box‐whisker plots represent the interquartile range, middle line indicates the median, and the whiskers indicate minimum/maximum values. (D–F) Correlation of mRNA expression levels of AATs (SLC1A5, SLC7A5, and SLC38A2) in CRC patient samples harboring KRAS wt (n = 317) and KRAS mutations (n = 217), retrieved from gene expression omnibus database (NCBI‐GEO). Error bars represent mean ± SEM; P‐values were calculated by unpaired t‐test.

Fig 3: Ras inhibition inhibits oligodendroglial cell morphological differentiation. (A,B) FBD-102b cells were treated with vehicle or Lanafamib (5 μM) and were allowed to be differentiated for 0 or 3 days. Differentiation efficiencies were divided into three categories and depicted in graphs (**, p < 0.01; n = 5 fields). The number of branches in cells is also counted and shown (**, p < 0.01; 50 cells). (C,D) Cells at 3 days following the induction of differentiation were collected, lysed, and immunoblotted with an antibody against PLP1, CNPase, SOX10, or actin. Immunoreactive band intensities were also compared to be depicted in graphs (**, p < 0.01; n = 3 blots).

Fig 4: Knockdown of ACE2 promotes the GTP-bound form of Ras in affinity-precipitation assay. (A) The lysates of siRNA-knocked down FBD-102b cells were used for affinity-precipitation (AP) assay, monitoring activated GTP-bound Ras with Ras-binding domain (RBD). Total Ras proteins were also shown. Band intensities were also compared to be depicted in graphs (**, p < 0.01; n = 3 blots). (B) The lysates of gRNA-knocked down cells were used for affinity-precipitation assay to monitor active GTP-bound Ras. Total Ras proteins were also shown. Band intensities were also compared to be depicted in graphs (**, p < 0.01; n = 3 blots).

Fig 5: SNARE interactions with KRAS are selectively SNORD50A/B-dependent. a Proximity ligation analysis (PLA) between KRAS and either SNAP23, SNAP29, or VAMP3, as well as single antibody controls in H23. Significance of WT cells determined by comparison to single antibody controls while significance of KO cells determined by comparison to WT. b Schematic of KRAS mutants used in proximity protein labeling experiments. c Strength of interaction between Raf1, SNAP23, SNAP29, or VAMP3 with KRAS mutants measured by proximity protein labeling followed by pulldown western blotting (n = 2) in H23. d Quantitation of c. e Interaction strength of Raf1 and SNAREs with KRASWT and KRASG13D in H23. f Quantification of e (n = 2). g Localization of HA-tagged KRAS and endogenous SNAP23, SNAP29, and VAMP3 visualized by super-resolution microscopy in A549. Scale bars are 5 µm. h Co-immunoprecipitation of endogenous RAS and SNARE proteins from A549 lysate. i Far western blot with spotted purified recombinant protein indicated by row name. Bound recombinant KRAS was detected with an anti-RAS antibody. Coomassie stain for total protein loading (middle) (n = 3, representative images shown). j Quantification of i. Error bars are s.e.m