Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: ALDH1A1 maintains the cancer stem-like cells properties of esophageal squamous cell carcinoma by activating the AKT signal pathway and interacting with β-catenin.

doi: 10.1016/j.biopha.2020.109940

Figure Lengend Snippet: Fig. 1. Differential expression of ALDH1A1 protein in ESCC tissues and the lost ALDH1A1 expression associated with poor overall survival. IHC analysis of ALDH1A1 expression in ESCC tissues. (A) Low expression of ALDH1A1 protein in hyperplastic esophagus tissues. (B, C) Low expression of ALDH1A1 protein in ESCC. (D) High expression of ALDH1A1 protein in ESCC. Scale bar =100 μm. (E) Overall survival of ESCC patients stratified in accordance with ALDH1A1 protein expression.

Article Snippet: The primary antibodies were incubated at 4 °C overnight: anti-AKT (dilution 1:1000), anti-AKT1 (1:1000), p-AKT (Ser473) (1:1000), p-AKT (Thr308) (1:1000) and Vimentin (1:500) were purchased from CST, βcatenin (1:2000), c-Myc (1:1000) from Proteintech, ALDH1A1 (1:1000, AL: ALDH1A1; L: low expression; H: High expression; HEs: Hyperplastic esophagus tissues; ESCCs: Esophageal squamous cell carcinoma tissues.

Techniques: Quantitative Proteomics, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: ALDH1A1 maintains the cancer stem-like cells properties of esophageal squamous cell carcinoma by activating the AKT signal pathway and interacting with β-catenin.

doi: 10.1016/j.biopha.2020.109940

Figure Lengend Snippet: Fig. 2. Ectopic expression of ALDH1A1 promoted spherogenesis and colony formation of ESCC cells in vitro. (A) Tumor sphere formation assays in KYSE-510/NC, KYSE-510/ALDH1A1, KYSE-150/NC, and KYSE-150/ALDH1A1 cells. Scale bar =200 μm. (B) Spherogenesis rate. The graph is the summarized data of three in- dependent experiments, ***P < 0.001. (C) ALDH1A1 expression promoted colony formation of KYSE-150 cells. The graph is the summarized data of three in- dependent experiments, ***P < 0.001. (D) Cell survival assay showed that ALDH1A1 expression promoted TE-1 cell resistance to 5-Fu. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies were incubated at 4 °C overnight: anti-AKT (dilution 1:1000), anti-AKT1 (1:1000), p-AKT (Ser473) (1:1000), p-AKT (Thr308) (1:1000) and Vimentin (1:500) were purchased from CST, βcatenin (1:2000), c-Myc (1:1000) from Proteintech, ALDH1A1 (1:1000, AL: ALDH1A1; L: low expression; H: High expression; HEs: Hyperplastic esophagus tissues; ESCCs: Esophageal squamous cell carcinoma tissues.

Techniques: Expressing, In Vitro, Clonogenic Cell Survival Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: ALDH1A1 maintains the cancer stem-like cells properties of esophageal squamous cell carcinoma by activating the AKT signal pathway and interacting with β-catenin.

doi: 10.1016/j.biopha.2020.109940

Figure Lengend Snippet: Fig. 3. Ectopic expression of ALDH1A1 promoted tumor initiation in vivo. (A) Equal KYSE-150/NC and KYSE-150/ALDH1A1 cells were injected into different axillaries (white arrow) of six BALB/c nude mice. (B) Transplanted tumor for- mation numbers.

Article Snippet: The primary antibodies were incubated at 4 °C overnight: anti-AKT (dilution 1:1000), anti-AKT1 (1:1000), p-AKT (Ser473) (1:1000), p-AKT (Thr308) (1:1000) and Vimentin (1:500) were purchased from CST, βcatenin (1:2000), c-Myc (1:1000) from Proteintech, ALDH1A1 (1:1000, AL: ALDH1A1; L: low expression; H: High expression; HEs: Hyperplastic esophagus tissues; ESCCs: Esophageal squamous cell carcinoma tissues.

Techniques: Expressing, In Vivo, Injection

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: ALDH1A1 maintains the cancer stem-like cells properties of esophageal squamous cell carcinoma by activating the AKT signal pathway and interacting with β-catenin.

doi: 10.1016/j.biopha.2020.109940

Figure Lengend Snippet: Fig. 4. ALDH1A1 expression regulated AKT and β-catenin expressions. (A) RT-qPCR assay detected mRNA expression levels of ALDH1A1, AKT1, β-catenin, Slug, c-Myc, and Vimentin in KYSE-150 and TE-1 cells. ** P < 0.01; *** P < 0.001; ns, not significant. (B, C) Western blot assay showed the ALDH1A1, p-AKT, total-AKT1, β-catenin, c-Myc, and β-actin protein expressions after NCT501 treatment or ALDH1A1 overexpression in ESCC cells, *: GAPDH. D, Western blot assay showed the ALDH1A1, AKT1, β-catenin, and β-actin expression levels after MK-2206 or NCT501 treatment. (E) Western blot gray scale for the relative protein levels of Fig. 4E. ***: P < 0.001.

Article Snippet: The primary antibodies were incubated at 4 °C overnight: anti-AKT (dilution 1:1000), anti-AKT1 (1:1000), p-AKT (Ser473) (1:1000), p-AKT (Thr308) (1:1000) and Vimentin (1:500) were purchased from CST, βcatenin (1:2000), c-Myc (1:1000) from Proteintech, ALDH1A1 (1:1000, AL: ALDH1A1; L: low expression; H: High expression; HEs: Hyperplastic esophagus tissues; ESCCs: Esophageal squamous cell carcinoma tissues.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Over Expression

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: ALDH1A1 maintains the cancer stem-like cells properties of esophageal squamous cell carcinoma by activating the AKT signal pathway and interacting with β-catenin.

doi: 10.1016/j.biopha.2020.109940

Figure Lengend Snippet: Fig. 5. ALDH1A1 interacted with β-catenin. Co-localization of ALDH1A1 and β-catenin in (A) adherent cells (Scale bar =20 μm) and (B) stem cell spheres (Scale bar =50 μm) of KYSE-510. Images were collected by a confocal microscope. (C) Co-IP, Western blot assay to detect the interaction of ALDH1A1 and β-catenin. (D) Western blot to detect β-catenin protein levels after cycloheximide (CHX) treatment. (E) Western blot to detect ALDH1A1and β-catenin protein expression in cytoplasm and nuclear of adherent KYSE-510 cells. (F) TOP/FOP flash activity assay revealed that ALDH1A1 promoted wnt/β-catenin activity in KYSE-293 T cells.

Article Snippet: The primary antibodies were incubated at 4 °C overnight: anti-AKT (dilution 1:1000), anti-AKT1 (1:1000), p-AKT (Ser473) (1:1000), p-AKT (Thr308) (1:1000) and Vimentin (1:500) were purchased from CST, βcatenin (1:2000), c-Myc (1:1000) from Proteintech, ALDH1A1 (1:1000, AL: ALDH1A1; L: low expression; H: High expression; HEs: Hyperplastic esophagus tissues; ESCCs: Esophageal squamous cell carcinoma tissues.

Techniques: Microscopy, Co-Immunoprecipitation Assay, Western Blot, Expressing, Activity Assay

Journal: Infectious agents and cancer

Article Title: Silencing of UBE2D1 inhibited cell migration in gastric cancer, decreasing ubiquitination of SMAD4.

doi: 10.1186/s13027-021-00402-2

Figure Lengend Snippet: Fig. 1 UBE2D1 was elevated in GC tissues. A The mRNA level of UBE2D1 was analyzed from TCGA-STAD data. B The mRNA level of UBE2D1 was detected in 25 pairs of tumors and paracancerous samples. ***, P < 0.001. C The protein level of UBE2D1 in tissue array (32 pairs) was determined by IHC staining. D Impact of UBE2D1 expression on OS in GC patients in TCGA

Article Snippet: Primary antibodies were listed as following: UBE2D1 (1:800, Proteintech, Rosemont, IL, USA), SMAD4 (1:1000, Abcam, St. Louis, MO, USA), MMP2 (1:1000, Abcam), MMP9 (1:800, Abcam), and β-actin (1:2000, Abcam).

Techniques: Immunohistochemistry, Expressing

Journal: Infectious agents and cancer

Article Title: Silencing of UBE2D1 inhibited cell migration in gastric cancer, decreasing ubiquitination of SMAD4.

doi: 10.1186/s13027-021-00402-2

Figure Lengend Snippet: Fig. 2 Expression level of UBE2D1 was measured by real-time PCR western blot assay. A UBE2D1 expression level in different cell lines, including GSE-1, AGS, BGC-823, MGC-803, MKN45, and SGC-7901. B Knock-down efficiency was evaluated after transduction with shUBE2D1 lentiviruses in AGS and MKN45 cells. ***, P < 0.001 versus shNC. C Overexpression efficiency was evaluated after transduction with UBE2D1 overexpression lentivirus in MGC-803 cells. ***, P < 0.001 versus vector

Article Snippet: Primary antibodies were listed as following: UBE2D1 (1:800, Proteintech, Rosemont, IL, USA), SMAD4 (1:1000, Abcam, St. Louis, MO, USA), MMP2 (1:1000, Abcam), MMP9 (1:800, Abcam), and β-actin (1:2000, Abcam).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Knockdown, Transduction, Over Expression, Plasmid Preparation

Journal: Infectious agents and cancer

Article Title: Silencing of UBE2D1 inhibited cell migration in gastric cancer, decreasing ubiquitination of SMAD4.

doi: 10.1186/s13027-021-00402-2

Figure Lengend Snippet: Fig. 4 Overexpression of UBE2D1 promoted cell migration in vitro. A–C Transwell (A and B) and wound healing (C) assays were performed to analyze cell migration after transduction with UBE2D1 overexpression lentivirus in MGC-803 cells. ***, P < 0.001 versus vector. D A western blot assay was performed to measure the protein levels of UBE2D1, MMP2, and MMP9

Article Snippet: Primary antibodies were listed as following: UBE2D1 (1:800, Proteintech, Rosemont, IL, USA), SMAD4 (1:1000, Abcam, St. Louis, MO, USA), MMP2 (1:1000, Abcam), MMP9 (1:800, Abcam), and β-actin (1:2000, Abcam).

Techniques: Over Expression, Migration, In Vitro, Transduction, Plasmid Preparation, Western Blot