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. 2024 Mar 23;8:74. doi: 10.1038/s41698-024-00574-1

Fig. 2. Inhibition of cell proliferation, migration, and invasion ability of NSCLC by knockdown of Rab27A.

Fig. 2

a The expression level of Rab27A in human NSCLC cells was detected by qRT-PCR and western blot. Data are shown as the mean ± SD of n = 3 technical replicates. b Rab27A mRNA and protein levels in Rab27A-knockdown NSCLC cells and negative control cells. c Rab27A mRNA and protein levels in Rab27A-overexpressing NSCLC cells and negative control cells. d CCK-8 assay to detect cell viability in A549 and H1299 cells; cell viability was determined at 24, 48, and 72 h. e Rab27A-knockdown inhibited the clonogenic ability of A549 and H1299 cells. f, g Cell lysates from A549 and H1299 cells infected with the indicated shRNAs were subjected to Western blot analysis to detect PCNA protein level. h, i Rab27A-knockdown inhibited the invasion and migration ability of A549 and H1299 cells. A549 and H1299 cells were allowed to migrate through an 8 μm pore Transwell. After 24 h, migrated cells were stained and counted in at least three microscopic fields (magnification, ×100). Then the cells were treated as above and allowed to invade through the Matrigel-coated membrane in Transwell. Invasive cells were stained and counted under a light microscope (magnification, ×100). j Rab27A-overexpression promoted the clonogenic ability of A549 and H1299 cells. k, l Cell lysates from A549 and H1299 cells with the indicated lentiviral vectors were subjected to Western blot analysis to detect PCNA protein level. m, n Rab27A-overexpression promoted the invasion and migration ability of A549 and H1299 cells. A549 and H1299 cells were allowed to migrate through an 8 μm pore Transwell. After 24 h, migrated cells were stained and counted in at least three microscopic fields (magnification, ×100). Then the cells were treated as above and allowed to invade through the Matrigel-coated membrane in Transwell. Invasive cells were stained and counted under a light microscope (magnification, ×100). o Rab27A-knockdown in A549 cells xenografts in nude mice (n = 6) at the experimental endpoint. Tumors were dissected and photographed as shown. p Tumor growth curves in mice (n = 6 in each group). q Each tumor formed was weighted. *P < 0.05; **P < 0.01; ***P < 0.001.