Figures & data
Figure 1. RASA4 downregulation in the CESC tissues. (a) The RASA4 mRNA levels in normal and neoplastic cervical tissues in the GEPIA database. (b) The RASA4 mRNA levels in normal and neoplastic cervical tissues in oncomine databases. (c) Kaplan–Meier curves for overall survival in CESC. (d) The ROC curve of RASA4 to assess the diagnostic value
Figure 2. RASA4 overexpression suppresses proliferation and colony formation in HeLa cells. (a) Western blotting demonstrating transient RASA4 overexpression in HeLa cells. RASA4 was exogenously overexpressed through cell transfection with the pLV-Flag-RASA4 vector. (b) RASA4 overexpression weakened the proliferation of HeLa cells. Cell proliferation was assessed through CCK8 assays at the indicated timepoints. Data are shown as mean ± SEM (n = 3). (c) RASA4 overexpression impaired the colony generation rate of HeLa cells. Cell colony formation was measured through colony formation assays. Data are shown as mean ± SEM (n = 3)
Figure 3. RASA4 depletion increases proliferation and colony formation in HeLa cells. (a) Western blotting demonstrated transient depletion of RASA4 in HeLa cells. RASA4 depletion was assessed using the CRISPR/Cas9 technology. (b) CCK8 assays demonstrated the impact of RASA4 deficiency on cell proliferation (n = 3). (c) Colony formation assays manifesting the impact of the RASA4 deficiency on the colony formation rate of HeLa cells
Figure 4. RASA4 depletion increases proliferation and colony formation in C-33A cells. (a) Western blotting demonstrated a transient depletion of RASA4 in the C-33A cells. RASA4 depletion was induced using the CRISPR/Cas9 technology. (b) CCK8 assays demonstrated the impact of RASA4 deficiency on cell proliferation (n = 3). (c) Colony formation assays manifested the impact of RASA4 deficiency on the colony formation rate of C-33A cells
Figure 5. RASA4 inactivated the HIF1α signaling pathway in CESC. (a) Dual-luciferase reporter assay demonstrated that the HIF1α-mediated luciferase activities decreased in HeLa cells infected with different doses of pLV-Flag vectors (0, 100, 200, 400 ng). (b) Western blotting revealed that survivin protein was reduced in the HeLa cells infected with different doses of pLV-Flag vectors (0, 100, 200, and 400 ng). (c) Detection of the HIF1α-mediated luciferase activity in RASA4-deficient HeLa cells. (d) Western blotting revealed that the survivin protein expression was increased in HeLa cells when RASA4 depleted. (e) RT-qPCR results showed that the escalated survivin mRNA levels by RASA4 depletion were partly reversed by LW6 treatment. (f) Western blotting results unveiled a gradual decrement of survivin protein in the RASA4-overexpressing HeLa cells along with an increasing dose of LW6 treatment. (g) Colony formation assays demonstrated that the LW6 treatment could offset the robust escalation of the colony formation number in RASA4-deficient HeLa cells
Figure 6. Knockout of RASA4 favors the growth of xenograft tumor in nude mice. (a) Pictures of tumor grown under RASA4 depletion; (b) The tumor volume growth curves from day 0 to day 16; (c) Tumor weight was analyzed after sacrificing the mice
