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. 2021 Aug 3;12(8):764. doi: 10.1038/s41419-021-04063-2

Fig. 2. ANLN-201 promotes SCC-9 cell proliferation, migration, and invasion.

Fig. 2

A ANLN was knocked out in SCC-9 cells using CRISPR/Cas9. B Cell counting kit-8 (CCK-8) was performed to measure cell proliferation in ANLN-sgRNA SCC-9 cells. *P < 0.05. C BrdU staining together with PI was performed to measure cell proliferation in ANLN-sgRNA SCC-9 cells. *P < 0.05, **P < 0.01. D Cell cycle analysis was used to analyze the change of cell cycles induced by ANLN knockout. *P < 0.05, **P < 0.01. E Transwell and transwell-matrigel assays were performed to measure cell migration and cell invasion caused by ANLN knockout. *P < 0.05, **P < 0.01. F Cell proliferation was assessed in ANLN-sgRNA SCC-9 cells transfected with GFP-ANLN-201 or GFP-ANLN-210. *P < 0.05, **P < 0.01. G Cell migration and cell invasion were analyzed in ANLN-sgRNA SCC-9 cells transfected with GFP-ANLN-201 or GFP-ANLN-210. *P < 0.05, **P < 0.01. H qRT-PCR was used to examine the mRNA level of ANLN-201 or ANLN-210 in ANLN-sgRNA SCC-9 cells transfected with ANLN overexpressing plasmids GFP-ANLN-201 or GFP-ANLN-210. *P < 0.05, **P < 0.01. I The immunofluorescence of ANLN-201 and ANLN-210 was assessed in ANLN-sgRNA SCC-9 cells transfected with ANLN overexpressing plasmids GFP-ANLN-201 or GFP-ANLN-210 using primary antibody against GFP. J The protein levels of ANLN-201 and ANLN-201 were estimated by immunoblot using the antibody against GFP.