Fig. 6
GOLGA7 is essential for the viability of NRAS-dependent cells and the transformation of oncogenic NRAS in mice. A Ba/F3-N cells (left panel) or Ba/F3-K cells (right panel) ± sgGOLGA7 were cultured without(w/o) IL-3. Cell viability of each cell line was measured by CellTiter-Glo Luminescent Cell Viability Assay 48 h after plating. Values are the means ± SD from three independent experiments, compared using two-sided Student’s t test. Ba/F3-N GOLGA7-sg1, ****p < 0.0001; Ba/F3-N GOLGA7-sg2, ****p < 0.0001; Ba/F3-K GOLGA7-sg1, ns p = 0.5134; Ba/F3-K GOLGA7-sg2, ns p = 0.2820. B Ba/F3-N cells (left panel) and Ba/F3-K cells (right panel) were cultured without(w/o) IL-3 for 30 min and analyzed subsequently by Western blotting using total or phospho-ERK, -AKT and -S6 antibodies. β-Actin was used as a loading control. C Cell viability of human cancer cell lines ± shGOLGA7/siGOLGA7 was detected by the CellTiter Glo assay. Values are the means ± SD from three independent experiments, compared using two-sided Student’s t test. Cell lines with mutant NRAS: THP-1(NRASG12D), ****p < 0.0001; OCI-AML3(NRASQ61L), ****p < 0.0001; HL-60(NRASQ61L), ****p < 0.0001; HepG2(NRASQ61L), ****p < 0.0001; SK-MEL-2(NRASQ61R), ****p < 0.0001. Cell lines with wild-type NRAS: MOLM-13(NRASWT), ns p = 0.4193; SK-MEL-30(NRASWT), ns p = 0.2299. Cell lines with mutant HRAS: T24(HRASG12V), ns p = 0.7229. Cell lines with mutant KRAS4B: NB4(KRAS4B.A18D), ns p = 0.1698. D Cells in (C) were analyzed by Western blotting using total or phospho-ERK, -AKT and -S6 antibodies. β-actin was used as a loading control. E The percentage of GFP-positive Ba/F3-N cells and the number of white blood cells in peripheral blood (PB) samples from mice with Ba/F3-N ± sgGOLGA7 cells injection after 10 days. ****p < 0.0001. E Kaplan–Meier survival curves of lethally irradiated mice with Ba/F3-N ± sgGOLGA7 cells injection. **** P < 0.0001. G The percentage of GFP-positive Ba/F3-K cells and the number of white blood cells in peripheral blood (PB) samples from mice with Ba/F3-K cells ± sgGOLGA7 injection after 10 days. ns p = 0.9853. H Kaplan–Meier survival curves of lethally irradiated mice with Ba/F3-K ± sgGOLGA7 cells injection, ns p = 0.9303