Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Hyperphosphorylation of ribosomal protein S6 predicts unfavorable clinical survival in non-small cell lung cancer

doi: 10.1186/s13046-015-0239-1

Figure Lengend Snippet: The upstream regulation signaling pathway of rpS6. a Separate overexpression of Akt1, Akt2 and Akt3 in HBE cells showed that only Akt2 (oe-Akt2) led to marked hyperphosphorylation of mTOR, p70S6K and rpS6 (left; All P < 0.05); while Akt1 or Akt3 alteration resulted in no detectable change of these proteins (left; All P > 0.05). In H1650 and SK-MES-1 cell lines, only the specific silence of Akt2 (sh-Akt2) caused significant loss of p-mTOR, p-p70S6K and p-rpS6 (middle and right; All P < 0.05), and no notable alteration with the Akt1 and Akt3 knockdown were found (sh-Akt1, sh-Akt3; middle and right; All P < 0.05). b H1650 and SK-MES-1 were treated with Akti-2 (Akt inhibitor XII) for 72 h in concentration of 0.8, 3.2 and 12.8 μM. Western blot assays revealed the strongest effect of 12.8 μM and the accompanied dephosphorylation of mTOR, p70S6K and rpS6 (All P < 0.05). oe-Akt1: overexpression of Akt1; oe-NCa1: negative control for oe-Akt1; oe-Akt2: overexpression of oe-Akt2; oe-NCa2: negative control for oe-Akt2; oe-Akt3: overexpression of oe-Akt3; oe-NCa3: negative control for oe-Akt3. sh-Akt1: knockdown for Akt1; sh-NCa1: negative control for sh-Akt1; sh-Akt2: knockdown for Akt2; sh-NCa2: negative control for sh-Akt2; sh-Akt3: knockdown for Akt3; sh-NCa3: negative control for sh-Akt3

Article Snippet: Specific protein antibodies included anti-total rpS6 (CST, #2217), anti-p-rpS6 (Ser235/236, CST, #4858), anti-p21 Cip1 (CST, #2947), anti-p27 Kip1 (CST, #3688), anti-CDK2 (CST, #2546), anti-CDK4 (CST, #12790), anti-Cyclin E (SAB, #29030), anti-cyclin D1 (CST, #2978), anti-p-Rb (Ser780, CST, #3590), anti-Paxillin (CST, #12065), anti-p-Paxillin (Tyr118; CST, #2541), anti-vimentin (CST, #5741), anti-N-cadherin (CST, #4061), anti-E-cadherin (CST, #3195), anti-MMP-9 (CST, #13667), anti-MMP-2 (CST, # 5741), anti-t-Akt1 (CST, #2967), anti-p-Akt1 (Ser473, CST, # 9018), anti-t-Akt2 (CST, #2964), anti-p-Akt2 (Ser474, CST, #8599), anti-t-Akt3 (Biorbyt, #orb6787), anti-p-Akt3 (Ser472, Biorbyt, #orb6790), anti-t-mTOR (CST, #2983), anti-p-mTOR (Ser2448, CST, #5536), anti-t-p70S6K (SAB, #21276) and anti-p-p70S6K (Ser424, SAB, #21276) and anti-β-actin (CST, #4970) as an internal control.

Techniques: Over Expression, Concentration Assay, Western Blot, De-Phosphorylation Assay, Negative Control

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: The mRNA expression analysis of MFSD12 and Its association with Clinical Features. (A) Differential expression analysis of MFSD12 between pan-cancer tissues and adjacent normal tissues in TCGA and GETx database. (B) Differential expression analysis of MFSD12 between tumor tissues and normal tissues in LIHC based on TCGA and GETx database. (C) Association of MFSD12 expression with clinical parameters in LIHC. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. MFSD12, Major Facilitator Superfamily Domain-containing 12; LIHC, liver hepatocellular carcinoma; TCGA, The Cancer Genome Atlas; GEO, Gene Expression Omnibus, AFP, Alpha-fetoprotein. "ns" stands for "not significant".

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Expressing, Quantitative Proteomics, Gene Expression

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: The protein expression analysis of MFSD12. (A) Pan-cancer protein expression profile of MFSD12 and representative IHC staining of tissue microarrays in HPA database. (B) IHC analysis of MFSD12 in LIHC tumor tissues and paired adjacent non-tumor liver tissues. (C) Quantification of immunostains for MFSD12 by IOD analysis. * P < 0.05, ** P < 0.01. IHC, immunohistochemistry; HPA, Human Protein Atlas; LIHC, liver hepatocellular carcinoma; IOD, integrated optical density;.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Expressing, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: Prognostic significance of MFSD12 expression across cancers and validation in LIHC Cohorts. (A) A pan-cancer Cox regression analysis was performed to assess MFSD12 expression. (B) OS analysis of MFSD12 in TCGA-LIHC data. (C) PFS analysis of MFSD12 in TCGA-LIHC data. (D) DFS analysis of MFSD12 in TCGA-LIHC data. (E) The prognostic significance of MFSD12 expression in LIHC patients was evaluated through both univariate and multivariate analyses. (F–H) Independent validation using external GEO cohorts corroborated the prognostic significance of MFSD12 in LIHC. AUC, Area Under Curve; CI, Confidence Interval; DFS, Disease-Free Survival; GEO, Gene Expression Omnibus; HR, Hazard Ratio; LIHC, Liver Hepatocellular Carcinoma; OS, Overall Survival; PFS, Progression-Free Survival; RFS, Relapse-Free Survival; ROC, Receiver Operating Characteristic; TCGA, The Cancer Genome Atlas; TPM, Transcripts Per Million.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Expressing, Biomarker Discovery, Gene Expression

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: Genomic alteration landscape of MFSD12 in LIHC. (A) Mutation spectrum of MFSD12 in pan-cancer analysis. (B) CNV analysis of MFSD12 in LIHC. (C) Genomic Landscape of mutations in MFSD12 within LIHC. (D) Classification profile of MFSD12 genetic variants in LIHC. (E) Comparative mutation profiling in MFSD12 low- and high-expressing subpopulations. (F) Protein-protein interaction network analysis of MFSD12 in LIHC. (G) Gene Co- Expression Network Correlated with MFSD12 Expression Patterns in LIHC. * P < 0.05, ** P < 0.01. CC, Cholangiocarcinoma; CNV, Copy Number Variation; LIHC, Liver Hepatocellular Carcinoma; MFSD12, Major Facilitator Superfamily Domain Containing 12; SNV, Single Nucleotide Variant; TCGA, The Cancer Genome Atlas; WES, Whole Exome Sequencing.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Mutagenesis, Expressing, Variant Assay, Sequencing

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: DNA methylation analysis of the MFSD12 genomic features in LIHC. (A) The chromosomal localization of MFSD12 within the human genome. (B) The genomic architecture of MFSD12 and its adjacent regions. (C) The dynamics of promoter methylation in LIHC and normal liver tissues. (D) MFSD12 methylation levels in tumor tissue samples compared to normal tissues. (E) Analysis of the correlation between MFSD12 expression and its methylation status. (F) The identification of tumor stage-specific methylation alterations. (G) The relationship between MFSD12 individual CpG site methylation values and CNV status (deep deletion, loss, neutral, gain, amplification). (H) The association of MFSD12 methylation with patient survival outcomes. *** P < 0.001. CpG, Cytosine-phosphate-Guanine dinucleotide; LIHC, Liver Hepatocellular Carcinoma; TCGA, The Cancer Genome Atlas; TNM, Tumor-Node-Metastasis staging system.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: DNA Methylation Assay, Methylation, Expressing, Amplification

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: MFSD12 functional enrichment analysis across immune-related pathways and biological processes in LIHC. (A) GO enrichment analysis of MFSD12-associated biological processes. (B) KEGG pathway enrichment of MFSD12. (C) The GSEA-GO enrichment profile of MFSD12 in the context of immune regulation, as indicated by the enrichment score. (D) The GSEA-KEGG enrichment profile of MFSD12 in the context of immune regulation, as indicated by the enrichment score. (E) Hallmark gene set enrichment of MFSD12 in LIHC. ES, Enrichment Score; GSEA, Gene Set Enrichment Analysis; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; MFSD12, Major Facilitator Superfamily Domain Containing 12.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Functional Assay

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: Integrative analysis of MFSD12 expression correlation with tumor microenvironment immunocytes in LIHC. (A) Correlation of MFSD12 with tumor microenvironment scores using algorithm of ESTIMATE database: association of MFSD12 with immune score, stromal score, and ESTIMATE score in LIHC. (B) Correlation of MFSD12 expression level with immune cell across 33 cancer types. (C) Comparison of immune cell proportions stratified by MFSD12 expression levels (Low vs. High) in TCGA_LIHC. (D) Relationship between MFSD12 expression and immune infiltration in LIHC, as analyzed by the ssGSEA algorithm. (E) Relationship between MFSD12 expression and immune infiltration in LIHC across a range of immune infiltration analysis tools and multiple genomic datasets. ** P < 0.01, *** P < 0.001. CIBERSORT, cell-type identification by estimating relative subsets of RNA Transcripts; Cor, Pearson correlation coefficient; ESTIMATE, estimation of stromal and immune cells in malignant tumor tissues using expression data; LIHC, Liver Hepatocellular Carcinoma; Pval, p-value; TCGA, The Cancer Genome Atlas; xCell, cell type enrichment analysis tool.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: Integrated analysis of the link between MFSD12 expression and immune-related genes. (A) The relationship between the expression levels of MFSD12 and Immune-related genes in pan-cancers; (B) The relationship between the MFSD12 expression levels and immune checkpoints in LUSC. (C) Relationship between MFSD12 expression and Immune-related genes in LIHC across a range of immune infiltration analysis tools and multiple genomic datasets. * P < 0.05, ** P < 0.01. LIHC, Liver Hepatocellular Carcinoma; Pearson, Pearson correlation coefficient; Cor, Correlation coefficient.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: Single-Cell analysis of MFSD12 in LIHC by scRNA-seq. (A) UMAP visualization of cell type distribution in LIHC. (B) UMAP expression profile of MFSD12 in LIHC. (C) Relative expression levels of MFSD12 across cell types. (D) UMAP visualization of cell distribution by location. (E) UMAP visualization of cell distribution by cancer subtype (Normal, HCC, CC). (F) Heatmap of G1/S and G2/M phase transition gene expression across cell types. (G) Cell number and proportion statistics for each cell type. (H) Expression Proportion of MFSD12 in Different Cell Types and Cancer Subtypes. UMAP, Uniform Manifold Approximation and Projection; MFSD12, Major Facilitator Superfamily Domain Containing 12; CD4T_conv, Conventional CD4+ T cells; CD8T_typical, Typical CD8+ T cells; CD8T_exhausted, Exhausted CD8+ T cells; T_prolif, Proliferating T cells; Treg, Regulatory T cells; NK_cell, Natural Killer cell; B_cell, B lymphocyte; Mono/Macro, Monocyte/Macrophage; HCC, Hepatocellular Carcinoma; CC, Cholangiocarcinoma; G1/S, G1/S phase transition genes; G2/M, G2/M phase transition genes.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Single-cell Analysis, Expressing, Sublimation, Gene Expression

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: A thorough analysis of the correlation between MFSD12 expression and drug response across various databases, as well as its association with survival outcomes. (A) Correlation between MFSD12 expression and drug resistance/sensitivity in the CTRP dataset. (B) Correlation between MFSD12 expression and drug resistance/sensitivity in the PRISM dataset. (C) Correlation between MFSD12 expression and drug resistance/sensitivity in the GDSC1 database. (D) Correlation between MFSD12 expression and drug resistance/sensitivity in the GDSC2 database. (E) Overall survival analysis of Hugo cohort 2016 (Anti-PD-1) and Nathanson cohort 2017 (Anti-CTLA-4). CTRIP, Cancer Therapeutics Response Portal; PRISM, Preclinical Repurposing of Medicines; GDSC1/GDSC2, Genomics of Drug Sensitivity in Cancer 1/2; Anti-PD-1, Anti-Programmed Cell Death Protein 1; Anti-CTLA-4, Anti-Cytotoxic T-Lymphocyte-Associated Protein 4; Log-rank, Log-rank test; Number at risk, Number of patients at risk at each time point.

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: MFSD12 promotes proliferation, metastasis and invasion of hepatocellular carcinoma cells and its potential correlation with HAVCR2/LGALS9 immune checkpoint axis

doi: 10.3389/fimmu.2025.1681887

Figure Lengend Snippet: The knockdown of MFSD12 inhibited the proliferation, migration, and invasion of LIHC cells, as well as the TIM-3/Galectin-9 signaling pathway. (A, B) RT-qPCR and Western blot validation of MFSD12 silencing efficiency using siRNAs (si-MFSD12–1 to −4) with GAPDH as loading control. (C) CCK-8 cell viability assay showing reduced HEP 3B2.1–7 cells proliferation after MFSD12 knockdown (si-MFSD12-3). (D) Transwell assay revealed a reduction in the migratory and invasive capabilities of HEP 3B2.1–7 cells following the knockdown of MFSD12. (E) Immunoblot analysis of EMT markers and TIM-3 axis components showing up-regulation of E-cadherin and down-regulation of Vimentin, MMP-2, MMP-9, HAVCR2 (TIM-3) and LGALS9 in si-MFSD12-treated cells. * P < 0.05, ** P < 0.01, *** P < 0.001. CTRL, control untreated; si-NC, negative control siRNA; si-MFSD12, MFSD12-targeting siRNA; E-cadherin, epithelial cadherin; MMP-2/9, matrix metalloproteinase-2/9; HAVCR2, hepatitis A virus cellular receptor 2 (TIM-3); LGALS9, lectin galactoside-binding soluble 9 (Galectin-9).

Article Snippet: The primary antibodies utilized in the Western blot analysis included: anti-rabbit MFSD12 (1:1000, Boster, China), anti-mouse GAPDH (1:30000, Proteintech, China), anti-rabbit E-cadherin (1:40000, Proteintech, China), anti-mouse Vimentin (1:40000, Proteintech, China), anti-rabbit MMP2 (1:1000, BIOSS, China), anti-rabbit MMP9 (1:1000, Affinity, China), anti-rabbit LGALS9 (1:1000, Abmat, China), and anti-rabbit HAVCR2 (1:1000, Boster, China).

Techniques: Knockdown, Migration, Quantitative RT-PCR, Western Blot, Biomarker Discovery, Control, CCK-8 Assay, Viability Assay, Transwell Assay, Negative Control, Virus, Binding Assay

Journal: Molecular cell

Article Title: Multisite phosphorylation of S6K1 directs a kinase phospho-code that determines substrate selection

doi: 10.1016/j.molcel.2018.11.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Full-length human S6K1 cDNA in pCMV6-Entry , Origene Technologies , Cat# RC217324.

Techniques: Recombinant, Purification, Mutagenesis, Cell Culture, Staining, Protease Inhibitor, Western Blot, Silver Staining, Transfection, Affinity Chromatography, Plasmid Preparation, Software