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. 2021 Mar 30;12(11):3098–3113. doi: 10.7150/jca.56340

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LOC promotes anchorage independent growth and cell invasive ability. A) (Top panel) Representative gel image shows multiplex PCR result of actin and LOC expression in LOC-overexpressing cells (LOC), control cells (Ctrl) and water as a negative control. (Middle panel) Number of colonies of LOC-overexpressing cells (LOC) and control (Ctrl) in soft agar. Colonies were stained with 1% (w/v) methyl green in methanol after incubating for 30 days post seeding. (Bottom panel) Representative figures taken from soft agar plates. B) (Top panel) Relative expression of LOC after knockdown in HepG2 cells (siLOC) compared to the knockdown control (siCtrl), as detected by qPCR. The expression is normalized against actin. (Middle panel) Number of colonies of LOC knockdown cells (siLOC) and control (siCtrl) in soft agar. Colonies were stained with 1%(w/v) methyl green in methanol after incubating for 28 days post seeding. (Bottom panel) Representative figures taken from soft agar plates. All data are shown as mean±SE of three biological replicates. **: P<0.01 compared with control. C) (Top panel) Representative gel image shows multiplex PCR result of actin and LOC expression in LOC-overexpressing cells (LOC), control cells (Ctrl) and water as a negative control. (Middle panel) Invasion profile of LOC-transfected cells (LOC) and control (Ctrl) using Transwell Matrigel assay. Cells were stained with Giemsa Stain solution before visualization under light microscope (x20 magnification). (Bottom panel) Representative pictures of invaded cells. ***: P<0.001 compared with control. D) (Top panel) Relative expression of LOC after knockdown in HepG2 cells (siLOC) compared to the knockdown control (siCtrl), as detected by qPCR. The expression is normalized against actin. (Middle panel) Invasion profile of LOC knockdown cells (siLOC) and control (siCtrl) using Transwell Matrigel assay. Cells were stained with Giemsa Stain solution before visualization under light microscope (x20 magnification). (Bottom panel) Representative pictures of invaded cells. All data are shown as mean±SE of three biological replicates. *:P<0.05 compared with control.