Twist1 Antibody - #AF4009

(E, F) Twist1 and Vimentin expression in the tumor tissues of Twi-L+control, Twi-L+circ-10720, Twi-H+siRNA control and TwiH+si-circ-10720 groups were analyzed via IHC.

Figure 2.| circ-10720 regulated by Twist1 exhibits oncogenic effects in HCC cells.(D) Western blot analysis of Twist1 and Cul2 protein levels in Twist1 overexpression or knockdown cells. Intensity of each band from biological triplicate experiments was quantified by densitometry with the Image J software with GAPDH as a normalizer. *P<0.05, **P<0.01, Student’s t-test.

Figure 4. |Intratumoral silencing of circ-10720 blocks the promotive effect of Twist1-mediated on Vimentin expression in a PDTX model of HCC. (E, F) Twist1 and Vimentin expression in the tumor tissues of Twi-L+control, Twi-L+circ-10720, Twi-H+siRNA control and TwiH+si-circ-10720 groups were analyzed via IHC. Each bar represents the mean ± SD for triplicate measurements of four xenografts in each group. **P<0.01, one-way ANOVA.

Fig. 3.|Sal reversed the drug resistance and inhibited HIF-1α signaling pathway. (I) Protein expression level of Twist1, Zeb1 and E-cadherin in PLC/PRF/5 cells treated hypoxia and hypoxia combined with Sal. Error bars represent the standard deviation of experiments performed in triplicate (*P b .05, **P b .01).

Fig. 5: Effect of NRF2 overexpression on the biological characteristics of Tu177/CDDPTNFAIP2-/- cells. A Quantitative PCR analysis shows the differences in NRF2 mRNA levels between Tu177/CDDP cells and their variants. B Immunofluorescence staining results display NRF2 protein expression in different cell lines, with red indicating NRF2 and blue indicating DAPI-stained nuclei, Bar = 25 μm. C Images of the sphere formation assay (top) and the corresponding statistical graphs of sphere numbers and diameters (bottom), comparing the sphere-forming abilities of the three cell lines, Bar = 100 μm. D, E Western blot analysis showing differences in EMT-related protein expression among the cell lines. F Transwell invasion assay (top, Bar = 50 μm) and wound healing assay (bottom, Bar = 100 μm) results demonstrate the invasion and migration abilities of the different cell lines. The number of invasive cells and the percentage of unhealed areas are compared among the three treatments; ① indicates Tu177/CDDP cells, ② indicates Tu177/CDDPTNFAIP2-/- cells, and ③ indicates Tu177/CDDPTNFAIP2-/-+oe-NRF2 cells. * represents p 

Fig. 2. Compounds SR‐3‐65 and WXM‐1‐170 regulated the protein level of EMT biomarkers and transcription factors in AGS and MGC803 cells. (A, B) SR-3–65 and WXM-1–170 altered the protein level of EMT biomarkers. AGS (A) and MGC803 (B) cells were treated with DMSO, indisulam, SR-3–65, or WXM-1–170 (10 µM) for 72 h. Cell lysates were immunoblotted. Mean ± SD (n = 3). One-way ANOVA, Dunnett’s post hoc analysis, *: P < 0.05, **: P < 0.01, ***: P < 0.001, ns: not significant. (C, D) SR-3–65 and WXM-1–170 downregulated the protein level of EMT-related transcription factors. AGS (C) and MGC803 (D) cells were treated with DMSO, indisulam, SR-3–65, or WXM-1–170 (10 µM) for 72 h. Cell lysates were immunoblotted. Mean ± SD (n = 3). One-way ANOVA, Dunnett’s post hoc analysis, *: P < 0.05, **: P < 0.01, ***: P < 0.001, ****: P < 0.0001, ns: not significant.

Fig. 4| Hsp90β stabilize Twist1 and promotes its deubiquitination by combining with USP19. a Hsp90β knockdown decreases Twist1 stability. SMMC-7721 cells were transfected with the Hsp90βsiRNA for 24h and then were treated with CHX (100mg/ml) at the indicated time points. b SMMC-7721 cells transfected with Hsp90βsiRNA were treated with 10µM MG132 and followed by immuno-blotting using anti-Twist1 and Hsp90β.

Fig. 6 | Hsp90β promotes tumor growth and VM formation relied on Twist1 in vivo and Hsp90 inhibitor depresses the promotion effect.e, f Hsp90β, Twist1, VE-cadherin, E-cadherin, Vimentin,MMP2, and MMP9 expression levels were measured in tumor tissue of the control, Hsp90β overexpression, Hsp90β+ Twist1 siRNA,control shRNA, and Hsp90β knockdown groups by IHC.

Fig. 6. XPO1 promotes sorafenib resistance by enhancing EMT. (A) GSEA enrichment score curve. (B) We used a microscope to observe the morphology of Huh/SR cells. (C) Western blot detection of EMT-related protein expression in Huh7 and Huh7/SR cells. Using Image J to analyze the band. Data are mean ± SD and analyzed by unpaired t test. n = 3. (D) Transwell assay of Huh7 and Huh7/SR cells. bar= 20 µm. n = 3. (E) IB analysis of WCL and anti-XPO1 IPs derived from Huh7 cells. IgG served as a negative control. (F) Molecular docking of XPO1 and Vimentin. (G) The subcellular localization of endogenous XPO1 and Vimentin was analyzed by fluorescence microscopy in Huh7/SR cells. The arrows highlight the nuclear colocalization of XPO1 and Vimentin by immunofluorescence staining. bar= 5 µm. n = 3. (H) R analysis of the correlation between XPO1 and VIM in TCGA HCC dataset. (I) Western blot detection of EMT-related protein expression. n = 3. (J) R analysis of the correlation between XPO1 and EMT markers in the TCGA HCC dataset.