Journal: Translational Oncology

Article Title: UCHL1 promotes temozolomide resistance in glioblastoma by inhibiting the ubiquitination-mediated degradation of keratin 8

doi: 10.1016/j.tranon.2026.102728

Figure Lengend Snippet: UCHL1 is positively associated with TMZ resistance in glioblastoma cells. (A) Cell viability curves for the U87MG, T98G, and U251 cell lines following TMZ treatment were evaluated using the CCK-8 assay. The IC50 values were determined through nonlinear regression analysis (curve fitting), with three replicates ( n = 3). (B) The IC50 values for TMZ-treated U251 cells were assessed after exposure to 100 nM MG132 (a proteasome inhibitor) or 5 μM CQ (an autophagy/lysosome inhibitor) ( n = 3). (C) Differential genes from the datasets GSE193957 , GSE211272 , GSE229600 , and GSE113510 were intersected with members of the ubiquitin proteasome family. Western blotting analysis (D) and RT-qPCR (E) were conducted to assess UCHL1 levels in the U87MG, T98G, and U251 cell lines ( n = 3). * p < 0.05, *** p < 0.001.

Article Snippet: The primary antibodies used included UCHL1 (Cat#13179, Cell Signaling Technology), KRT8 (Cat#AG2433, Beyotime and Cat# bsm-52419R, Bioss, Beijing, China), PARP (Cat#9542, Cell Signaling Technology), Myc Tag (Cat#2276, Cell Signaling Technology), DYKDDDDK Tag (Cat#14793, Cell Signaling Technology), HA Tag (Cat#3724, Cell Signaling Technology), GAPDH (Cat#60004-1-Ig, Proteintech, Wuhan, China), γ-H2AX (Cat#9718, Cell Signaling Technology) and Bcl-2 (Cat#3498, Cell Signaling Technology).

Techniques: CCK-8 Assay, Ubiquitin Proteomics, Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: UCHL1 promotes temozolomide resistance in glioblastoma by inhibiting the ubiquitination-mediated degradation of keratin 8

doi: 10.1016/j.tranon.2026.102728

Figure Lengend Snippet: Expression of UCHL1 and its clinical significance in glioma patients. (A) UCHL1 expression was evaluated in a tissue microarray of glioma samples. Representative images show varying levels of UCHL1 in TMZ-sensitive and TMZ-resistant tissues. (B) Survival probability was determined according to UCHL1 expression in primary and recurrent glioma samples using the CGGA dataset. (C) UCHL1 expression in WHO grade II, III, and IV tissues from the CGGA dataset.

Article Snippet: The primary antibodies used included UCHL1 (Cat#13179, Cell Signaling Technology), KRT8 (Cat#AG2433, Beyotime and Cat# bsm-52419R, Bioss, Beijing, China), PARP (Cat#9542, Cell Signaling Technology), Myc Tag (Cat#2276, Cell Signaling Technology), DYKDDDDK Tag (Cat#14793, Cell Signaling Technology), HA Tag (Cat#3724, Cell Signaling Technology), GAPDH (Cat#60004-1-Ig, Proteintech, Wuhan, China), γ-H2AX (Cat#9718, Cell Signaling Technology) and Bcl-2 (Cat#3498, Cell Signaling Technology).

Techniques: Expressing, Microarray

Journal: Translational Oncology

Article Title: UCHL1 promotes temozolomide resistance in glioblastoma by inhibiting the ubiquitination-mediated degradation of keratin 8

doi: 10.1016/j.tranon.2026.102728

Figure Lengend Snippet: Dual effects of UCHL1 knockdown and overexpression on TMZ sensitivity. (A) Western blotting and (B) RT-qPCR analyses of UCHL1 expression in U251 cells following UCHL1 knockdown ( n = 3). (C) Western blotting and (D) RT-qPCR analyses of UCHL1 expression in U87MG cells transfected with either an empty vector or a UCHL1-containing lentivirus ( n = 3). (E) Cell viability curves for U251 cells with control shRNA (shNC) and UCHL1 shRNA (sh UCHL1 ) after TMZ treatment. (F) Cell viability curves for U87MG cells transfected with either the empty vector or the UCHL1 overexpression vector following TMZ exposure. (G) Western blotting analysis showing Bcl-2 and cPARP expression in U251 cells with UCHL1 knockdown after TMZ treatment ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, ns means no significance.

Article Snippet: The primary antibodies used included UCHL1 (Cat#13179, Cell Signaling Technology), KRT8 (Cat#AG2433, Beyotime and Cat# bsm-52419R, Bioss, Beijing, China), PARP (Cat#9542, Cell Signaling Technology), Myc Tag (Cat#2276, Cell Signaling Technology), DYKDDDDK Tag (Cat#14793, Cell Signaling Technology), HA Tag (Cat#3724, Cell Signaling Technology), GAPDH (Cat#60004-1-Ig, Proteintech, Wuhan, China), γ-H2AX (Cat#9718, Cell Signaling Technology) and Bcl-2 (Cat#3498, Cell Signaling Technology).

Techniques: Knockdown, Over Expression, Western Blot, Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Control, shRNA

Journal: Translational Oncology

Article Title: UCHL1 promotes temozolomide resistance in glioblastoma by inhibiting the ubiquitination-mediated degradation of keratin 8

doi: 10.1016/j.tranon.2026.102728

Figure Lengend Snippet: UCHL1 induces TMZ resistance by deubiquitination-mediated stabilization of KRT8 in glioblastoma cells. (A) We identified chemoresistant and anti-apoptotic genes in the database, as revealed by IP-MS. Western blotting analysis shows KRT8 levels in glioblastoma cells subjected to UCHL1 knockdown (B) and UCHL1 overexpression (C). (D) The Co-IP assay indicates the interaction between UCHL1 and KRT8 in U87MG cells. (E) HEK293T cells were co-transfected with the indicated plasmids, harvested 48 h post-transfection, and the ubiquitination level of Myc‑tagged KRT8 was determined. (F) HEK293T cells were co-transfected with the indicated plasmids, harvested 48 h post-transfection, and subjected to ubiquitination assay to specifically assess the contribution of K27‑linked ubiquitin chains to KRT8 ubiquitination. * p < 0.05.

Article Snippet: The primary antibodies used included UCHL1 (Cat#13179, Cell Signaling Technology), KRT8 (Cat#AG2433, Beyotime and Cat# bsm-52419R, Bioss, Beijing, China), PARP (Cat#9542, Cell Signaling Technology), Myc Tag (Cat#2276, Cell Signaling Technology), DYKDDDDK Tag (Cat#14793, Cell Signaling Technology), HA Tag (Cat#3724, Cell Signaling Technology), GAPDH (Cat#60004-1-Ig, Proteintech, Wuhan, China), γ-H2AX (Cat#9718, Cell Signaling Technology) and Bcl-2 (Cat#3498, Cell Signaling Technology).

Techniques: Protein-Protein interactions, Western Blot, Knockdown, Over Expression, Co-Immunoprecipitation Assay, Transfection, Ubiquitin Proteomics

Journal: Translational Oncology

Article Title: UCHL1 promotes temozolomide resistance in glioblastoma by inhibiting the ubiquitination-mediated degradation of keratin 8

doi: 10.1016/j.tranon.2026.102728

Figure Lengend Snippet: KRT8 is a key protein in UCHL1-mediated TMZ resistance in glioblastoma. (A) Survival probability was assessed based on KRT8 expression levels in primary and recurrent glioma samples using data from the CGGA dataset. (B) KRT8 expression levels in WHO grade II, III, and IV glioma tissues, as derived from the CGGA dataset. (C) Western blotting analysis depicting KRT8 expression in U251-sh UCHL1 cells following KRT8 overexpression. (D) Cell viability curves for KRT8 overexpression in U251-sh UCHL1 cells subjected to TMZ treatment. (E) Survival analysis based on UCHL1 and KRT8 co‑expression in glioblastoma patients (TCGA dataset). *** p < 0.001.

Article Snippet: The primary antibodies used included UCHL1 (Cat#13179, Cell Signaling Technology), KRT8 (Cat#AG2433, Beyotime and Cat# bsm-52419R, Bioss, Beijing, China), PARP (Cat#9542, Cell Signaling Technology), Myc Tag (Cat#2276, Cell Signaling Technology), DYKDDDDK Tag (Cat#14793, Cell Signaling Technology), HA Tag (Cat#3724, Cell Signaling Technology), GAPDH (Cat#60004-1-Ig, Proteintech, Wuhan, China), γ-H2AX (Cat#9718, Cell Signaling Technology) and Bcl-2 (Cat#3498, Cell Signaling Technology).

Techniques: Expressing, Derivative Assay, Western Blot, Over Expression