Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: High expression of RAB40C was associated with a poor prognosis in HCC patients. A The expression of RAB40C in HCC tissues was compared with those in matched non-carcinoma tissues in the TCGA datasets. B , C RAB40C expression in HCC samples with different stages and grades was analyzed using UALCAN database. D RAB40C mRNA level was examined by qPCR in 17 pairs of HCC tissues and matched non-carcinoma hepatic tissues. E , F RAB40C protein level in pairs of HCC tissues and matched non-carcinoma hepatic tissues. The intensity of RAB40C protein was analyzed by Image J software. G , H The Overall survival and disease-free survival of HCC patients were analyzed by using GEPIA database. Survival curves were plotted by the Kaplan-Meier method and analyzed by the log-rank test. All *** P < 0.001

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Software

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: RAB40C promoted the growth, migration and tumorigenesis of HCC cells. A , B , C The growth-promoting effect of RAB40C in HCC cells was examined by CCK8 and crystal violet assays. D , E , F The inhibitory effect of knockdown RAB40C on the growth of HCC cells was detected by CCK8 and crystal violet assay. G The effects of RAB40C overexpression on HCC cell migration was assessed by transwell assays. H , I Overexpression of RAB40C increased the volume and weight of tumors formed by HepG2 cells. J The effects of RAB40C knockdown on HCC cell migration was assessed by transwell assays. K , L Knockdown of RAB40C attenuated the volume and weight of tumors formed by Huh7 cells. All * P < 0.05, ** P < 0.01

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Migration, Knockdown, Crystal Violet Assay, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: RAB40C was positively correlated with EGFR expression and downstream signaling in HCC cells. A GSEA enrichment plot for the EGFR signaling gene modules in RAB40C-high or RAB40C-low groups. B The top 20 enrichment of differentially expressed genes by GO assay. C Western Blot analyses of RAB40C and EGFR expression in human HCC tissues. D Correlation between RAB40C and EGFR expression in HCC patients. E , F Western blot analyses of EGFR and the signaling proteins in HCC cells with high or low expression of RAB40C. G , H The mRNA levels of RAB40C and EGFR were detected by qPCR in HCC cells with overexpression-RAB40C or RAB40C-knockdown. I , J Protein stability of EGFR was determined

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Western Blot, Over Expression, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: RAB40C maintained EGFR stability through K63-linked ubiquitination. A , B , C Co-immunoprecipitation showed interaction between RAB40C and EGFR. D The ubiquitination level of EGFR was detected in RAB40C-knockdown cells. E , F The K48-linked or K63-linked ubiquitination of EGFR was detected in RAB40C-knockdown cells by western blot. G The ubiquitination level of EGFR was detected in RAB40C-overexpression cells. H , I The K48-linked or K63-linked ubiquitination of EGFR was detected in RAB40C-overexpression cells by western blot

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Ubiquitin Proteomics, Immunoprecipitation, Knockdown, Western Blot, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: RAB40C promoted the ubiquitination of EGFR via the E3 ligase TRIM21. A Silver-stained gel showing differential bands between control and RAB40C-overexpressing samples. B , C , D The interaction between RAB40C, TRIM21 and EGFR. E HEK293T cells were co-transfected with Tag-Myc-TRIM21 and HA-Ub/K48-only or HA-Ub/K63 only followed by IP analysis. F The K63-linked ubiquitination of EGFR was reduced after attenuated RAB40C expression in the presence of TRIM21. G The K63-linked ubiquitination of EGFR was reduced after TRIM21 was weakened

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Ubiquitin Proteomics, Staining, Control, Transfection, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: STAT3 increased the transcription level of RAB40C by binding to the promoter. A Diagram of predicting STAT3 binding to the RAB40C promoter. B , C STAT3 regulated the mRNA level of RAB40C. D , E The effect of STAT3 expression on RAB40C protein level was detected by western blot. F The RAB40C protein level was examined after Stattic treatment. G Luciferases experimental diagram of STAT3 targeting binding to the RAB40C promoter. H , I , J The luciferase and ChIP assays were performed to examine the regulation of STAT3 on RAB40C transcription in 293 T cells, IgG served as the negative control. K The inhibitory effect of STAT3 inhibitor stattic on the colony formation and transwell of HepG2 cells. Al * P < 0.05, ** P < 0.01, *** P < 0.001, ns: not significant

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Binding Assay, Expressing, Western Blot, Luciferase, Negative Control

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: Artemisinin, a ligand of RAB40C, displayed a promising potential in HCC therapy. A A 2D model of binding of artemisinin to amino acid residues of RAB40C. B 3D model of RAB40C’s optimal binding mechanism in the protein pocket (artemisinin depicted as colored sticks). C Amino acid residues of RAB40C interacting with the artemisinin in 3D (color sticks). D , E The inhibition effect of artemisinin on EGFR was detected by western blot. F The k63-linked ubiquitination of EGFR by artemisinin was detected by IP analysis. G , H The inhibitory effect of artemisinin on cell viability of HepG2 and Huh7 cells by colony formation assay

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Binding Assay, Inhibition, Western Blot, Ubiquitin Proteomics, Colony Assay

Journal: Cell Communication and Signaling : CCS

Article Title: RAB40C recruiting TRIM21 facilitates the progression of hepatocellular carcinoma by stabilizing EGFR

doi: 10.1186/s12964-025-02580-7

Figure Lengend Snippet: Schematic representation of the molecular mechanism that RAB40C facilitates the progression of hepatocellular carcinoma through recruiting TRIM21 to stabilize EGFR by K63 ubiquitination

Article Snippet: Anti-RAB40C (sc-514826) antibody was purchased from Santa Cruz Biotechnology.

Techniques: Ubiquitin Proteomics

Journal: Journal of Bacteriology

Article Title: Analyzing the Regulatory Role of the HigA Antitoxin within Mycobacterium tuberculosis

doi: 10.1128/jb.00454-10

Figure Lengend Snippet: FIG. 5. Recombinant HigA protein directly and specifically binds to the higB P2 promoter. Protein-DNA binding assays were performed between pure refolded HigA-His protein and the 68-bp 32P-labeled higB P2 promoter probe (23 fmol) using increasing amounts of protein (A), unlabeled higB P2 promoter fragment (identical to the probe) as specific competitor (B), unlabeled higB P1 promoter fragment as nonspecific competitor (C), and anti-HigA antibody (D). The concentrations of recombinant protein used were 3.5, 7, 14, and 28 M for lanes 2 to 5, respectively, 0 M for lanes 1 and 17, and 56 M for all other lanes. Competitor DNA fragments were added in 5-, 10-, 100-, and 200-fold molar excess of the labeled probe (lanes 8 to 11 and 13 to 16). Anti-HigA antibody was used at final dilutions of 1 in 5,000, 500, 50, and 5 for lanes 19 to 22, respectively.

Article Snippet: For competition or supershift assays, the appropriate amount of unlabeled competitor DNA or rabbit anti-HigA antibody (raised and purified by Cambridge Research Biochemicals Ltd. against a synthesized peptide equivalent to residues 112 to 129 of the HigA protein) was added to the reaction mixture.

Techniques: Recombinant, Binding Assay, Labeling

Journal: Journal of Bacteriology

Article Title: Analyzing the Regulatory Role of the HigA Antitoxin within Mycobacterium tuberculosis

doi: 10.1128/jb.00454-10

Figure Lengend Snippet: FIG. 6. Identification of the HigA DNA-binding motif and the effect of mutating this motif on HigA binding and higB P2 promoter activity. (A) The wild-type and mutated sequences of the 68-bp higB P2 promoter fragment. The experimentally determined P2 promoter transcriptional start site (45) and putative 10 and 35 sequences are in boldface and boxed, respectively. The identified perfect palindromic DNA motif is underlined, with mutated bases highlighted in gray. (B) DNA-binding assays were performed between pure refolded HigA-His protein (0 and 56 M for lanes marked and , respectively) and the 32P-labeled mutated higB P2 promoter probe (23 fmol). (C) Protein-DNA-binding assays were performed between pure refolded HigA-His protein (56 M) and the 32P-labeled wild-type higB P2 promoter probe (23 fmol) using increasing amounts of unlabeled mutated higB P2 promoter fragment as competitor (5-, 10-, 100-, and 200-fold molar excess of the labeled probe for lanes 2 to 5, respectively). (D) -Gal assays were performed on cell-free protein extracts of wild-type M. tuberculosis H37Rv and the higB-Rv1957 deletion strain containing the promoter-lacZ fusion plasmids pASF32 (wild-type higB P2 promoter) and pASF58 (mutated higB P2 promoter) alongside the empty vector control (pEJ414). The data presented are averages of the results for three biological replicates, each assayed in duplicate, and error bars represent standard deviations. The -Gal activity level for the higB-Rv1957 deletion strain containing pASF32 was significantly higher than the equivalent wild-type-strain level (, P 0.05; Student’s t test). The -Gal activity level for the higB-Rv1957 deletion strain containing pASF58 was not significantly different from the equivalent wild-type-strain level (P 0.05; Student’s t test).

Article Snippet: For competition or supershift assays, the appropriate amount of unlabeled competitor DNA or rabbit anti-HigA antibody (raised and purified by Cambridge Research Biochemicals Ltd. against a synthesized peptide equivalent to residues 112 to 129 of the HigA protein) was added to the reaction mixture.

Techniques: Binding Assay, Activity Assay, Labeling, Plasmid Preparation, Control

Journal: Journal of Bacteriology

Article Title: Analyzing the Regulatory Role of the HigA Antitoxin within Mycobacterium tuberculosis

doi: 10.1128/jb.00454-10

Figure Lengend Snippet: FIG. 7. HigA does not bind to the next-most-closely related motif within the genome. (A) Sequence of the 68-bp fragment containing the DNA motif (underlined) between whiB5 and Rv0023, with the two bases mismatched to the HigA DNA-binding motif highlighted in gray. (B) Protein- DNA binding assays were performed between pure refolded HigA-His protein (0 and 56 M for lanes marked and , respectively) and the 32P-labeled whiB5-Rv0023 probe (23 fmol). (C) Protein-DNA-binding assays were performed between pure refolded HigA-His protein (56 M) and the 32P-labeled wild-type higB P2 promoter probe (23 fmol) using increasing amounts of unlabeled whiB5-Rv0023 fragment as competitor (5-, 10-, 100-, and 200-fold molar excess of the labeled probe for lanes 2 to 5, respectively).

Article Snippet: For competition or supershift assays, the appropriate amount of unlabeled competitor DNA or rabbit anti-HigA antibody (raised and purified by Cambridge Research Biochemicals Ltd. against a synthesized peptide equivalent to residues 112 to 129 of the HigA protein) was added to the reaction mixture.

Techniques: Sequencing, Binding Assay, Labeling