Fig 1: DLL1 overexpression neutralized the effect of FBXW7 overexpression on α-SMA and Collagen Ⅰ expressions and Notch signaling pathway in LX-2 cells. LX-2 cells were transfected with or without FBXW7/DLL1 silencing or overexpression plasmid. The expression levels of α-SMA and Collagen Ⅰ in LX-2 cells were measured by qRT-PCR (a) and western blot (b and c), where GAPDH was employed as the internal control. The protein levels of NOTCH2, NOTCH3, and HES1 in LX-2 cells were determined by western blot (d and e), with GAPDH serving as the internal control. The significant difference was measured by ANOVA, followed by Tukey’s post hoc test. ** p < 0.001 vs NC; ^^ p < 0.001 vs DLL1; ++ p < 0.001 vs FBXW7. ANOVA, analysis of variance; DLL1, delta-like ligand 1; FBXW7, F-box and WD repeat domain containing 7; α-SMA, α-smooth muscle actin; NC, negative control; qRT-PCR, quantitative reverse transcription polymerase chain reaction.
Fig 2: DLL1 overexpression offset the effects of FBXW7 overexpression on LX-2 cell cycle and proliferation. LX-2 cells were subjected to transfection of FBXW7/DLL1 silencing or overexpression plasmid. Colony formation assay was used to detect cell proliferation (a and b). Flow cytometry was employed to analyze cell cycle (c and d). The significant difference was detected by ANOVA, followed by Tukey’s post hoc test. ** p < 0.001 vs NC; ^^ p < 0.001 vs DLL1; ++ p < 0.001 vs FBXW7. ANOVA, analysis of variance; DLL1, delta-like ligand 1; FBXW7, F-box and WD repeat domain containing 7; NC, negative control.
Fig 3: Effects of FBXW7 on cell cycle and the levels of α-SMA and Collagen Ⅰ in LX-2 cells. LX-2 cells were transfected with or without FBXW7 silencing or overexpression plasmid. A cell cycle kit and flow cytometry were applied for the determination of cell cycle (a and b). The expressions of α-SMA and Collagen Ⅰ in LX-2 cells were measured by qRT-PCR (c) and western blot (d and e), where GAPDH was used as the internal control. The significant difference was determined by ANOVA, followed by Tukey’s post hoc test. ** p < 0.01 vs NC; ^^ p < 0.01 vs shNC. ANOVA, analysis of variance; FBXW7, F-box and WD repeat domain containing 7; shFBXW7, short hairpin RNA against FBXW7; shNC, shRNA negative control; qRT-PCR, quantitative reverse transcription polymerase chain reaction.
Fig 4: Effects of FBXW7 on LX-2 cell viability and proliferation. LX-2 cells were transfected with or without FBXW7 silencing or overexpression plasmid. The expression of FBXW7 in the transfected cells was determined by qRT-PCR and western blot, with GAPDH serving as the internal control (a, b and c). LX-2 cell viability at 0, 24, 48, and 72 h was analyzed by CCK-8 assay (d). Colony formation assay was used to detect the colony number of LX-2 cells (e and f). The significant difference was measured by ANOVA, followed by Tukey’s post hoc test. ** p < 0.001 vs NC; ^^ p < 0.001 vs shNC. ANOVA, analysis of variance; CCK-8, cell counting kit-8; FBXW7, F-box and WD repeat domain containing 7; h, hour; shFBXW7, short hairpin RNA against FBXW7; shNC, shRNA negative control; qRT-PCR, quantitative reverse transcription polymerase chain reaction.
Fig 5: DLL1 overexpression reversed the effect of FBXW7 overexpression on LX-2 cell viability. LX-2 cells were transfected with or without FBXW7/DLL1 silencing or overexpression plasmid. Western blot was applied to detect DLL1 and FBXW7 protein levels in LX-2 cells (a–d). CCK-8 assay was performed to detect LX-2 cell viability (e). The significant difference was determined by ANOVA, followed by Tukey’s post hoc test. ** p < 0.001 vs NC; ^^ p < 0.001 vs DLL1; ++ p < 0.001 vs FBXW7. CCK-8, cell counting kit-8; DLL1, delta-like ligand 1; FBXW7, F-box and WD repeat domain containing 7; NC, negative control.
Supplier Page from Sino Biological, Inc. for Human FBXW7 transcript variant 3 Gene ORF cDNA clone expression plasmid