Fig 2: Rap1GAP counteracts the effect of miR-3121-3p inhibitor. (A,B) The ability of tumor migration in TPC-1, BCPAP, and K1 cells (upper panel in A,B) decreases with miR-3121-3p inhibitors and elevates after transfection with Rap1GAP small interfering RNA. Invasion assay shows the same result (lower panel in A). (C,D) Student’s t-test clarified the significance. INVS indicates invasion assays. *, P<0.05; **, P<0.01; ***, P<0.001. Scale bar in A, 40 µm; scale bar in B, 80 µm.

Fig 3: SOX9 regulates the invasion and migration of A549 cells by activating the RAP1 signaling pathway. A-B, RT-qPCR to test the expression levels of SOX9 and RAP1 in cells of each group; C-D, Western blot to detect the expression levels of RAP1, RAP1GAP and RasGRP3 in cells; E, Transwell to observe cell invasion levels; F, Scratch migration assay to observe the migration ability of cells. **P < 0.01, vs. sh-NC; ##P < 0.01, vs. sh-SOX9; ¡÷¡÷P < 0.01, vs. RAP1.

Fig 4: Rap1GAP overexpression aggravates cardiomyocyte hypertrophy and further inhibits Ang II-induced autophagy. (a) The structure of the construct of expression plasmid. (b) mRNA levels of Rap1GAP, ANF, and BNP in Rap1GAP overexpressing and intact cardiomyocytes determined using RT-PCR. (c) Representative images and quantification from α-actinin staining showing cardiomyocyte surface area in Rap1GAP overexpressing and intact cardiomyocytes. Scale bar: 20 μm; n = 10. (d) Western blot analysis for LC3BII/I, p62, and cleaved-caspase-3/caspase-3 in Rap1GAP overexpressing and intact cardiomyocytes. (e) Representative images and quantifications from TEM showing autophagy lysosomes, autophagosomes, and mitochondria. The red arrows indicate autophagy lysosome or autophagosome, and the yellow boxes indicate mitochondria. Scale bar: 1.0 μm. ∗P < 0.05 and ∗∗P < 0.01. Results are the mean ± SEM of three independent experiments (n = 3).

Fig 5: Rap1GAP overexpression aggravates Ang II-induced ROS production and apoptosis. (a) Phosphorylated protein and total protein expression of AMPK, AKT, mTOR, and p70s6K measured using Western blot analysis. (b) ROS production in Rap1GAP overexpressing and control cardiomyocytes evaluated using DHE fluorescent dyes. Scale bar: 50 μm. (c) Cell apoptosis determined by TUNEL staining. Scale bar: 100 μm. ∗P < 0.05 and ∗∗P < 0.01. Data are the mean ± SEM of three independent experiments (n = 3).