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. 2022 Feb 23;12:774988. doi: 10.3389/fonc.2022.774988

Figure 4.

Figure 4

Tspan9 promotes in vitro OS cell migration and invasion. (A) GFP expression was indicative of stable lentiviral transduction of HOS cells with shTspan9 (shTspan9#1 or shTspan9#2) or shNC constructs at near 100% efficiency (fluorescent microscopy). Tspan9 knockdown efficiency was also confirmed via qRT-PCR (lower left panel) and Western blotting (lower right panel). (B) OS cell migration was assessed in a wound-healing assay using cells stably expressing shTspan9 or shNC. (C) The impact of Tspan9 knockdown on OS cell migration and invasion was assessed via a Transwell approach. (D) GFP expression was indicative of successful Tspan9 overexpression, as confirmed via qRT-PCR and Western blotting relative to Mock control. Wound-healing (E) and Transwell assays (F, G) were conducted to assess the impact of Tspan9 on the migratory and invasive activity of OS cells. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; Student’s t-test. GFP, green fluorescent protein; sh, short hairpin RNA.