Skip to main content
. 2021 May 17;10(5):310–320. doi: 10.1302/2046-3758.105.BJR-2020-0262.R3

Fig. 1.

Fig. 1

G protein subunit alpha Q (GNAQ) regulates tumour growth through mitogen-activated protein kinase (MAPK) signalling but does not increase apoptosis levels. a) GNAQ transcript levels in A549, A549_shNC, and A549_shGNAQ cells were measured by semiquantitative polymerase chain reaction (PCR) (left), and GNAQ protein expression was visualized by western blotting (right). b) Cell proliferation capacities in GNAQ-knockdown cells. c) GNAQ small interfering RNA (siRNA) was transfected into lung cancer cell line (A549, NCI-H1299, and HCC827), with negative siRNA serving as a control (upper panel). GNAQ transcript levels in cells were measured by semi-quantitative PCR (lower panel). d) Analysis of tumour growth for four weeks after injection of A549_shNC and A549_shGNAQ cells into the calvarial bone. Schematic diagram of location and the number of injected cells in mouse (left). Photographs show size comparisons of xenograft tumours in the different groups (n = 5 per group) (middle). A tumour growth curve was plotted based on the tumour volumes and the days elapsed (right). e) Representative images showing nodules on the lung surface and lung histology in mice. Mice (n = 5 per group) were injected via the tail vein with A549_shNC and A549_shGNAQ cells. Three months later, the mice were euthanized, and tumour nodules in the lungs were assessed. Two mice in the control group (A549_shNC) died before being euthanized. Lung nodules were visualized by haematoxylin and eosin (H&E) staining, magnification ×40 (Scanscope XT, Aperio, Leica Biosystems, USA). f) The expression of GNAQ and phosphylated extracellular signal-regulated kinase (p-ERK) in each group is shown by immunohistochemical staining of the xenograft tumours, magnification ×40 (Scanscope XT). g) The phosphorylation status of ERK1/2 and signal transducer and activator of transcription 3 (STAT3) was determined by western blotting. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; shNC, negative control short hairpin RNA; siNC, negative control small interfering RNA.