Journal: International journal of medical sciences

Article Title: Ube2S regulates Wnt/β-catenin signaling and promotes the progression of non-small cell lung cancer.

doi: 10.7150/ijms.40243

Figure Lengend Snippet: Figure 1. Immunostaining of Ube2S in normal lung tissues and NSCLC tissues. Bronchial epithelial cells (black arrow) and alveolar cells (grey arrow) show negative immunostaining for Ube2S (A). Bronchial epithelial cells (black arrow) and submucosal glands (grey arrow) show negative immunostaining for Ube2S (B). Weak and focal immunostaining of Ube2S is evident in some bronchial epithelial cells (C) and submucosal glands (D). Strong immunostaining of Ube2S is present in the cytoplasm and nuclei of squamous cell carcinoma (E) and adenocarcinoma (F) cells. (A: ×100; B, D, F: ×200; C, E: ×400)

Article Snippet: Ube2S cDNA clone was purchased from Origene (Rockville, MD, USA).

Techniques: Immunostaining

Journal: International journal of medical sciences

Article Title: Ube2S regulates Wnt/β-catenin signaling and promotes the progression of non-small cell lung cancer.

doi: 10.7150/ijms.40243

Figure Lengend Snippet: Figure 2. Survival function. Ube2S expression in non-small cell lung cancer is significantly associated with poor patient survival (34.9 ± 5.5 versus 56.4 ± 7.3 months) (Log rank test, p < 0.05)

Article Snippet: Ube2S cDNA clone was purchased from Origene (Rockville, MD, USA).

Techniques: Expressing

Journal: International journal of medical sciences

Article Title: Ube2S regulates Wnt/β-catenin signaling and promotes the progression of non-small cell lung cancer.

doi: 10.7150/ijms.40243

Figure Lengend Snippet: Figure 3. The function of Ube2S in lung cancer cells. Western blots show that Ube2S is expressed in bronchial epithelial HBE cells, and lung cancer A549 and NCI-H1299 cells (A). The MTT assay shows that overexpression of Ube2S in A549 cells significantly promotes cancer cell proliferation (B) (p < 0.05). The wound scratch healing assay shows that overexpression of Ube2S in A549 cells significantly promotes cancer cell migration (C) (*p < 0.05)

Article Snippet: Ube2S cDNA clone was purchased from Origene (Rockville, MD, USA).

Techniques: Western Blot, MTT Assay, Over Expression, Migration

Journal: International journal of medical sciences

Article Title: Ube2S regulates Wnt/β-catenin signaling and promotes the progression of non-small cell lung cancer.

doi: 10.7150/ijms.40243

Figure Lengend Snippet: Figure 4. Ube2S regulates Wnt/β-catenin signaling molecules and activity. Western blots show that overexpression of Ube2S in A549 cells significantly upregulates Wnt/β-catenin signaling molecules including β-catenin, cyclin D1, and MMP7 (A) (p < 0.05). The luciferase assay shows that overexpression of Ube2S in A549 cells significantly upregulates Wnt/β-catenin signaling activity (B) (p < 0.05)

Article Snippet: Ube2S cDNA clone was purchased from Origene (Rockville, MD, USA).

Techniques: Activity Assay, Western Blot, Over Expression, Luciferase

Journal: International journal of medical sciences

Article Title: Ube2S regulates Wnt/β-catenin signaling and promotes the progression of non-small cell lung cancer.

doi: 10.7150/ijms.40243

Figure Lengend Snippet: Figure 5. Wnt/β-catenin signaling inhibitor abolished the function of Ube2s in lung cancer cells. The MTT assay shows that addition of the Wnt/β-catenin signaling inhibitor, ETC-159, significantly inhibits the ability of Ube2S to promote cancer cell proliferation

Article Snippet: Ube2S cDNA clone was purchased from Origene (Rockville, MD, USA).

Techniques: MTT Assay

Journal: bioRxiv

Article Title: Unbiased CRISPR Synthetic Lethal Screening for Genetic Vulnerabilities in Succinate Dehydrogenase (SDH)-loss Model of Paraganglioma

doi: 10.1101/2025.08.26.672429

Figure Lengend Snippet: (A) Schematic illustration of the regulatory relationship between UBE2M and UBE2F in regulating apoptosis. (B) Absolute growth rates of Sdhb +/+ and Sdhb -/- imCCs following knockdown with non-targeting control (shNT), shUBE2M, or shUBE2F. UBE2M knockdown selectively enhances growth in Sdhb -/- cells, while UBE2F knockdown strongly suppresses growth in both genotypes. P-values are calculated using Bonferroni correction for multiple t-tests and the number of asterisks indicates degree of significance. (C) Relative growth rates from (B) normalized to shNT controls, highlighting the lineage-specific increase in proliferation upon UBE2M knockdown in Sdhb -/- cells. Error bars account for propagation of error. (D) Western blot analysis of ubiquitylated NOXA (high-molecular-weight smear, ∼40–250 kDa), CUL5, UBE2F, UBE2M, GAPDH (loading control), and SDHB in Sdhb +/+ and Sdhb -/- imCCs after knockdown with shNT, shUBE2M, or shUBE2F. Protein depletion was observed only 8-10 cell doublings after treatment. The pattern suggests altered CUL5-associated substrate ubiquitylation upon UBE2F loss and stabilization of UBE2F substrates upon UBE2M depletion in the Sdhb -/- background.

Article Snippet: Primary antibodies used were: α-CRISPR-Cas9 (Abcam #ab210571; 1:1000); α-SDHB (Abcam #459230; 1:2000), α-GAPDH (Abcam #ab8245; 1:5000), α-UBE2M (Abcam #ab109507; 1:5000), α-UBE2F (Proteintech #17056-1-AP; 1:5000), α-CUL5 (Abcam #ab184177; 1.5:5000), α-NEDD8 (Invitrogen #PA5-17476; 1.5:5000) and α-NOXA (Boster bio #M02287; 1:1000).

Techniques: Knockdown, Control, Western Blot, High Molecular Weight

Journal: bioRxiv

Article Title: Unbiased CRISPR Synthetic Lethal Screening for Genetic Vulnerabilities in Succinate Dehydrogenase (SDH)-loss Model of Paraganglioma

doi: 10.1101/2025.08.26.672429

Figure Lengend Snippet: (A) Schematic figure of the neddylation cascade and pharmacological targeting at the E1 (NAE1) or E2 (UBE2F) levels. Drugs were used to block UBE2F (UBE2Fi) or NEDD8-activating enzyme (NAEi). (B) Dose–response data showing relative growth rates (treated/untreated) of Sdhb +/+ and Sdhb -/- cells following treatment with the UBE2F inhibitor (UBE2Fi) for 2 doublings. (C) Immunoblot analysis of CUL5, UBE2F, UBE2M, GAPDH, and SDHB after UBE2Fi treatment for 0, 6, 12, and 24 h, showing progressive reduction in CUL5 neddylation and changes in UBE2F protein levels. (D) Dose– response data showing relative growth rates after treatment with the NEDD8-activating enzyme inhibitor (NAEi) for 72 h. Both Sdhb +/+ and Sdhb -/- cells exhibit growth suppression, with Sdhb -/- cells showing a stronger effect. (E) Immunoblot analysis of NEDD8, UBE2M, UBE2F, GAPDH, and SDHB after MLN4924 treatment for 0, 6, 12, and 24 h, confirming global inhibition of neddylation through suppression of NEDD8 and no changes in E2 enzyme levels.

Article Snippet: Primary antibodies used were: α-CRISPR-Cas9 (Abcam #ab210571; 1:1000); α-SDHB (Abcam #459230; 1:2000), α-GAPDH (Abcam #ab8245; 1:5000), α-UBE2M (Abcam #ab109507; 1:5000), α-UBE2F (Proteintech #17056-1-AP; 1:5000), α-CUL5 (Abcam #ab184177; 1.5:5000), α-NEDD8 (Invitrogen #PA5-17476; 1.5:5000) and α-NOXA (Boster bio #M02287; 1:1000).

Techniques: Blocking Assay, Western Blot, Inhibition