Figure 6
Elevated FAM83A expression is dependent on MEK/ERK signalling-driven JUNB transcription. (a) FAM83A mRNA expression induced upon serum stimulation of HPAC cells as assessed by qRT-PCR at the indicated times. (b) HPAC, Panc04.03 and Panc05.04 cells were pretreated with MEK and ERK pathway inhibitors prior to 4 h of serum stimulation to assess for induced FAM83A expression. (c) HPAC cells transfected with siRNA smartpools targeting EGR1, FOSB, JUNB, KRAS, FAM83A and a non-targeting (NT) control were assessed for serum-induced FAM83A expression. The individual knockdowns are shown on the left and identified as E (EGR1), F (FOSB), J (JUNB) and K (KRAS). FAM83A expression in each knockdown group is shown on the right together with the FAM83A knockdown (si83A). Student’s t-test was used to assess statistical significance with ** representing p < 0.01. (d–e) HPAC cells infected with lentivirus encoding two different shRNAs targeting JUNB (shJUNB1, shJUNB2) and control (shGFP) were assessed for serum-induced FAM83A expression (d) and growth (e). HPAC cells infected with lentivirus encoding shRNAs targeting FOSB (shFOSB) and control (shGFP) were assessed for FOSB and serum-induced FAM83A expression (f). All data are representative of three independent experiments.
