Fig. 2
From: KRAS regulation by small non-coding RNAs and SNARE proteins
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
