Fig 1: Ectopic FBP1 suppressed Snail-induced EMT and tumour growth in SMMC-7721 cells.a Western blot analysis showed expression changes of E-cadherin and Vimentin induced by Snail expression were hindered by forced expression of FBP1 in SMMC-7721 cells. Ectopic FBP1 expression did not significantly affect Snail, Slug, ZEB1 and Twist1 expression. b Immunofluorescence assay showed SMMC-7721-snail-FBP1 cells expressed higher E-cadherin but lower Vimentin levels and appeared more like epithelial cells than SMMC-7721-Snail cells. The magnifications used were ×200. c Transwell migration analyses showed FBP1 expression significantly inhibited cell migration induced by Snail overexpression in SMMC-7721 cells. d Representative images of day 42 tumours in mice transplanted with SMMC-7721, SMMC-7721-Snail and SMMC-7721-Snail-FBP1. e, f The mean tumour diameters and weights in each group are shown. g The representative images of Snail, FBP1 and E-cadherin expression in transplanted tumours. The magnifications used were ×200. *P < 0.05, NS not significant, compared with control. The data presented in a–c are based on three independent repeats. The number of mice for each group in d is 9. MW molecular weight
Fig 2: FGD5-AS1 attenuated the inhibitory effects of miR-493-5p on the malignant phenotypes of NSCLC cellsA. Expression of miR-493-5p in NSCLC cells transfected with miR-493-5p mimics or co-transfected with FGD5-AS1 overexpression plasmid was detected by qRT-PCR.B-D. CCK-8 assay (B), scratch healing assay (C) and Transwell assays (D) were employed for detecting proliferation, migration and invasion of the transfected cells.E and F. Western blot was used for quantifying the expressions of E-cadherin, N-cadherin, Snail, Twist1, ZEB1 and Vimentin protein in transfected cells. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 3: Knockdown of FGD5-AS1 repressed the proliferation, migration, invasion and EMT of NSCLC cellsA. qRT-PCR was employed for detecting the expression of FGD5-AS1 in H1650 and A549 cells after transfection of siRNA.B. CCK-8 method was used for detecting NSCLC cell proliferation after FGD5-AS1 was silenced.C and D. Scratch healing assay was employed for examining NSCLC cell migration after FGD5-AS1 was silenced.E and F. Invasion of NSCLC cells was examined with Transwell assay after FGD5-AS1 was silenced.G and H. Western blot was used for quantifying the expressions of E-cadherin, N-cadherin, Snail, Twist1, ZEB1 and Vimentin proteins in NSCLC cells after transfection. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 4: miR-149 repressed osteogenic differentiation by reducing Twist1 degradation through AKT1.A The expression of AKT1 and Twist1 in periosteal tissues determined by western blot analysis. B IHC staining analysis of STRO-1 protein in periosteal tissues (scale bar = 25 µm) and the quantitative analysis. C Periosteum calcified nodule staining (scale bar = 50 µm). D ALP activity in periosteal tissues. E OCN content in periosteal tissues. *p < 0.05 compared with rats receiving the Masquelet-induced membrane technique treated with inhibitor-NC, #p < 0.05 compared with rats receiving the Masquelet-induced membrane technique treated with miR-149-inhibitor + vector. Data were expressed as mean ± standard deviation. Comparison among multiple groups was conducted by one-way ANOVA, followed by Tukey’s post hoc test. MI the Masquelet-induced membrane technique, NC negative control, IHC immunohistochemistry, STRO-1 stromal cell antigen 1, ALP alkaline phosphatase, OCN osteocalcin, ANOVA one-way analysis of variance.
Fig 5: AKT1 facilitated osteogenic differentiation of MSCs by promoting Twist1 degradation.A The phosphorylation level of Twist1 and Twist1 protein level determined by western blot analysis, *p < 0.05 compared with sham-operated rats or control cells. B The phosphorylation level of Twist1 and Twist1 protein level determined by western blot analysis, *p < 0.05 compared with cells transfected with vector, #p < 0.05 compared with cells transfected with si-vector. C The expression of Twist1 measured by western blot analysis. D The expression of AKT1 and Twist1 determined by western blot analysis. E ALP activity in cells. F OCN content in cells. G Calcium deposition determined by calcified nodule staining (scale bar = 25 µm). *p < 0.05 compared with cells transfected with vector, #p < 0.05 compared with cells transfected with AKT1 + vector, &p < 0.05 compared with cells transfected with AKT1 + si-vector, n = 3. H The expression of AKT1 and Twist1 determined by western blot analysis. *p < 0.05 compared with cells transfected with mimic-NC + vector, #p < 0.05 compared with cells transfected with mimic-miR-149 + vector. Data were expressed as mean ± standard deviation. Unpaired t test was used for data comparison between two groups and one-way ANOVA was used for data comparison among multiple groups, followed by Tukey’s post hoc test. MSC mesenchymal stem cell, ALP alkaline phosphatase, OCN osteocalcin, ANOVA one-way analysis of variance.
Supplier Page from Abcam for Anti-Twist (phospho S68) antibody [EP18178]