a, Targeting positions of sgRNAs and antisense oligonucleotides
(ASOs) in the EMICERI and MOB3B locus.
b, Expression of EMICERI and its neighboring
genes in A375 cells transduced with non-targeting (NT) or
EMICERI-targeting sgRNAs and dCas9. c,
Schematic for bimodal perturbation of EMICERI transcription.
sgRNAs 1-3 use MS2 loops to recruit MS2-P65-HSF1 to dCas9 to activate
EMICERI, whereas sgRNAs 4-5 recruit only dCas9 to repress
EMICERI. d, Correlation between
MOB3B and EMICERI expression produced by
different combinations of sgRNAs with and without MS2 loops. e,
Schematic for inserting polyadenylation signals (pAS) downstream of the
EMICERI TSS. SV40, Simian virus 40; PGK, phosphoglycerate
kinase. f,
EMICERI expression after SAM activation of
EMICERI for the wild type and pAS clones. g,
MOB3B expression after the same perturbations as (f).
h, Vemurafenib resistance after SAM activation of
EMICERI. i, Expression of
EMICERI and MOB3B after ASO knockdown of
EMICERI in the context of SAM activation. j,
Expression of MOB3B and EMICERI after ASO
knockdown of MOB3B in the context of SAM activation. All values
are mean ± SEM with n = 4.
****P < 0.0001;
***P < 0.001;
**P < 0.01;
*P < 0.05. ns = not
significant.