Journal: Cell death & disease

Article Title: MEX3A promotes the malignant progression of ovarian cancer by regulating intron retention in TIMELESS.

doi: 10.1038/s41419-022-05000-7

Figure Lengend Snippet: Fig. 2 MEX3A knockdown inhibits the cell proliferation and tumor growth of ovarian cancer cells in vitro and in vivo. A, B MEX3A knockdown and overexpression efficiency were confirmed by qRT-PCR (n = 4 biologically independent samples) and western blotting. C Growth curve showing the growth of ovarian cancer cells upon MEX3A knockdown or overexpression (n = 5 biologically independent samples). D EdU assay showed the proliferation ability changes of ovarian cancer cells after MEX3A knockdown (n = 5 biologically independent samples). E The clonogenic assay showed the clonogenic capacity changes of ovarian cancer cells after MEX3A knockdown and overexpression (n = 5 biologically independent samples). F Images of tumors isolated from subcutaneous implantation of nude mice with MEX3A knockdown and control cells (n = 6 mice per group). G Tumor size and weight statistics. P-value was obtained by unpaired t-test. Results represent the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01.

Article Snippet: The MEX3A overexpression lentiviral plasmid was purchased from OriGene (RC215359L3, USA).

Techniques: Knockdown, In Vitro, In Vivo, Over Expression, Quantitative RT-PCR, Western Blot, EdU Assay, Clonogenic Assay, Isolation, Control

Journal: Cell death & disease

Article Title: MEX3A promotes the malignant progression of ovarian cancer by regulating intron retention in TIMELESS.

doi: 10.1038/s41419-022-05000-7

Figure Lengend Snippet: Fig. 3 MEX3A downregulating suppresses the metastasis of ovarian cancer cells. A Transwell assays were used to evaluate the effect of MEX3A expression on the migration of ovarian cancer cells (n = 5 biologically independent samples). B Transwell assays were used to evaluate the effect of MEX3A expression on the invasion of ovarian cancer cells (n = 5 biologically independent samples). P-value was obtained by unpaired t-test. Results represent the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01.

Article Snippet: The MEX3A overexpression lentiviral plasmid was purchased from OriGene (RC215359L3, USA).

Techniques: Expressing, Migration

Journal: Cell death & disease

Article Title: MEX3A promotes the malignant progression of ovarian cancer by regulating intron retention in TIMELESS.

doi: 10.1038/s41419-022-05000-7

Figure Lengend Snippet: Fig. 6 TIMELESS knockdown reversed the biological effects caused by MEX3A overexpression. A TIMELESS knockdown reduced the colony formation abilities of MEX3A-overexpressing HEY cells (n = 5 biologically independent samples). B Xenograft experiments showed TIMELESS knockdown partially reversed the effect of MEX3A overexpression on the growth of ovarian cancer cell in vivo (n = 6 mice per group). C, D TIMELESS inhibition partially reversed the effect of MEX3A overexpression on cell migration and invasion (n = 5 biologically independent samples). P-value was obtained by unpaired t-test. Results represent the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01.

Article Snippet: The MEX3A overexpression lentiviral plasmid was purchased from OriGene (RC215359L3, USA).

Techniques: Knockdown, Over Expression, In Vivo, Inhibition, Migration

Journal: Cell death & disease

Article Title: MEX3A promotes the malignant progression of ovarian cancer by regulating intron retention in TIMELESS.

doi: 10.1038/s41419-022-05000-7

Figure Lengend Snippet: Fig. 8 Model showing the effect of MEX3A/TIMELESS signaling on the malignancy of ovarian cancer cells. Abundant MEX3A regulates TIMELESS expression and performs oncogenic functions through maintaining the normal splicing of TIMELESS. At the deficiency of MEX3A, TIMELESS splicing goes wrong which leads to reduction of TIMELESS and decreases the oncogenic ability of MEX3A.

Article Snippet: The MEX3A overexpression lentiviral plasmid was purchased from OriGene (RC215359L3, USA).

Techniques: Expressing

Journal: The EMBO Journal

Article Title: The RNA-binding E3 ubiquitin ligase MEX-3C links ubiquitination with MHC-I mRNA degradation

doi: 10.1038/emboj.2012.218

Figure Lengend Snippet: MEX-3C specifically regulates HLA-A but not HLA-B or HLA-C expression in primary human NK cells and modulates NK cell killing. ( A ) Immunoblot analysis of MEX-3C from freshly isolated and activated primary human NK cells. NK cells were purified from healthy blood donors and activated for 1–3 weeks. MEX-3C siRNA nucleofection results in efficient depletion in NK cells at 72 h post-nucleofection. ( B ) Immunoblot analysis of MEX-3C in the NK-92 (upper panel) and NKL cell lines (lower panel), following stimulation with IL-2 (350 U/ml) at the different time points shown. ( C ) Immunoblot analysis of MEX-3C downregulation in NKL cells after 72 h post-nucleofection. ( D ) Depletion of MEX-3C in NKL cells results in an allotype-specific increase of surface HLA-A2. Cytofluorometric analysis of HLA-A2, -B, -C, and TfR levels in mock (dashed line) and siMEX-3C cells (solid line). Isotype control (solid grey). ( E ) MEX-3C depletion increases HLA-A2 but not HLA-B mRNA levels in NKL cells. HLA-A2 , pan-HLA-B , and MEX-3C mRNA levels were measured by qRT–PCR in mock and siMEX-3C nucleofected cells. Quantitative RT–PCR analysis of MEX-3C siRNA knockdown cells showed a significant 2.03±0.56-fold increase in HLA-A2 mRNA levels over mock cells ( P -value=0.010, n =3). Transcript levels are relative to GAPDH and normalized to mock levels, set as 1 and expressed as the mean±s.d. of three independent experiments ( P -value=0.01). ( F ) Depletion of MEX-3C increases HLA-A2 mRNA half-life. Mock (dashed line) or siMEX-3C (solid line) treated NKL cells were incubated with Actinomycin D (5 μg/ml). HLA-A2 and HLA-B mRNA levels were measured by qRT–PCR and normalized against HPRT . Results are the mean±s.d. of three independent experiments. ( G ) Depletion of MEX-3C in primary human NK cells results in an allotype-specific increase of surface HLA-A (HLA-A2 and -A3) but not HLA-B or -C. Cytofluorometric analysis of HLA-A2, -A3, -B, and -C levels in mock (dashed line) and siMEX-3C nucleofected cells (solid line). Isotype control (solid grey). ( H ) Reduced target cell killing in MEX-3C depleted primary NK cells. 51 Cr release assay (percent specific lysis) using mock siRNA (dashed line) or siMEX-3C (solid line) nucleofected NK cells against K562 target cells. Error bars show the standard error of the six replicates at each point. The results are representative of three independent experiments using polyclonal NK cells from unrelated donors.

Article Snippet: Primary antibodies used were rabbit polyclonal anti-RKHD2 (MEX-3C) antibody (Pro-Sci Incorporated) used at 1:1500 dilution and incubated overnight in PBST containing 5% milk.

Techniques: Expressing, Western Blot, Isolation, Purification, Control, Quantitative RT-PCR, Knockdown, Incubation, Release Assay, Lysis

Journal: Scientific Reports

Article Title: Comprehensive bioinformatics analysis of MEX3 family genes in hepatocellular carcinoma

doi: 10.1038/s41598-025-02057-0

Figure Lengend Snippet: Expressions, receiver operator characteristic (ROC) curves and survival analysis of MEX3 family genes in hepatocellular carcinoma (HCC). (* p < 0.05, ** p < 0.01, *** p < 0.001). ( A ) The expression of MEX3A, MEX3B, MX3C and MEX3D in HCC tissue and normal tissue. ( B ) The (ROC) curves of MEX3 family members of HCC. ( C-J ) Survival analysis of MEX3 family genes in HCC patients. The overall survival (OS) and disease-specific survival (DSS) survival curves comparing patients with high and low MEX3 family genes expression in HCC.

Article Snippet: Membranes were blocked at room temperature with 5% non-fat milk for 2 h, then incubated overnight at 4 °C with primary antibodies: anti-MEX3A(1:800, CBG122A, CUSABIO) and anti-β-actin (1:2000, 20536-1-AP, Proteintech), MEX3B(1:1000, SA240318 × 09, ORIGEN), MEX3C(1:1000, 15I0053, affinity), MEX3D(1:1000, BE01081275, BIOSS), Signal detection was performed using an ECL chemiluminescence system (Epizyme).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Comprehensive bioinformatics analysis of MEX3 family genes in hepatocellular carcinoma

doi: 10.1038/s41598-025-02057-0

Figure Lengend Snippet: Further validating the expression of MEX3A, MEX3B, MX3C and MEX3D in HCC and normal control. (* p < 0.05, ** p < 0.01, *** p < 0.001). ( A ) The GSE36376 dataset from the GEO database validated the expression levels of MEX3A, MEX3B, MEX3C, and MEX3D were significantly elevated in HCC tissues compared to normal controls. ( B ) qRT-PCR and WB to explore the mRNA (Fig. 2B) and protein levels (Fig. 2C).of MEX3 family genes in HCC cells.

Article Snippet: Membranes were blocked at room temperature with 5% non-fat milk for 2 h, then incubated overnight at 4 °C with primary antibodies: anti-MEX3A(1:800, CBG122A, CUSABIO) and anti-β-actin (1:2000, 20536-1-AP, Proteintech), MEX3B(1:1000, SA240318 × 09, ORIGEN), MEX3C(1:1000, 15I0053, affinity), MEX3D(1:1000, BE01081275, BIOSS), Signal detection was performed using an ECL chemiluminescence system (Epizyme).

Techniques: Expressing, Control, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Comprehensive bioinformatics analysis of MEX3 family genes in hepatocellular carcinoma

doi: 10.1038/s41598-025-02057-0

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) of the MEX3 family in HCC. Pathways were considered significantly enriched when they met the criteria: false discovery rate (FDR) < 0.25, P-value < 0.05, and normalized enrichment score (NES) exceeding 1. (A) GSEA for MEX3A based on Reactome pathways. ( B ) GSEA for MEX3B based on Reactome pathways. ( C ) GSEA for MEX3C based on Reactome pathways. ( D ) GSEA for MEX3D based on Reactome pathways.

Article Snippet: Membranes were blocked at room temperature with 5% non-fat milk for 2 h, then incubated overnight at 4 °C with primary antibodies: anti-MEX3A(1:800, CBG122A, CUSABIO) and anti-β-actin (1:2000, 20536-1-AP, Proteintech), MEX3B(1:1000, SA240318 × 09, ORIGEN), MEX3C(1:1000, 15I0053, affinity), MEX3D(1:1000, BE01081275, BIOSS), Signal detection was performed using an ECL chemiluminescence system (Epizyme).

Techniques:

Journal: Scientific Reports

Article Title: Comprehensive bioinformatics analysis of MEX3 family genes in hepatocellular carcinoma

doi: 10.1038/s41598-025-02057-0

Figure Lengend Snippet: MEX3 family genes expressions and tumor immunity. In the bar graph, MEX3A ( A ), MEX3B ( B ), MEX3C ( C ) and MEX3D ( D ) expressions were related to 24 immune infiltration cells of HCC. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Membranes were blocked at room temperature with 5% non-fat milk for 2 h, then incubated overnight at 4 °C with primary antibodies: anti-MEX3A(1:800, CBG122A, CUSABIO) and anti-β-actin (1:2000, 20536-1-AP, Proteintech), MEX3B(1:1000, SA240318 × 09, ORIGEN), MEX3C(1:1000, 15I0053, affinity), MEX3D(1:1000, BE01081275, BIOSS), Signal detection was performed using an ECL chemiluminescence system (Epizyme).

Techniques:

Journal: Scientific Reports

Article Title: Comprehensive bioinformatics analysis of MEX3 family genes in hepatocellular carcinoma

doi: 10.1038/s41598-025-02057-0

Figure Lengend Snippet: The relationships between immune checkpoint genes, including PD-1 (PDCD1), PD-L1 (CD274), and CTLA4, and MEX3 family genes of HCC. ( A-C ) The relationships between PDCD1 ( A ), CD274 ( B ), CTLA4 ( C ) and MEX3A in HCC. ( D-F ) The relationships between PDCD1 ( D ), CD274 ( E ), CTLA4 ( F ) and MEX3B in HCC. ( G-I ) The relationships between PDCD1 ( G ), CD274 ( H ), CTLA4 ( I ) and MEX3C in HCC. ( J-L ) The relationships between PDCD1 ( J ), CD274 ( K ), CTLA4 ( L ) and MEX3D in HCC.

Article Snippet: Membranes were blocked at room temperature with 5% non-fat milk for 2 h, then incubated overnight at 4 °C with primary antibodies: anti-MEX3A(1:800, CBG122A, CUSABIO) and anti-β-actin (1:2000, 20536-1-AP, Proteintech), MEX3B(1:1000, SA240318 × 09, ORIGEN), MEX3C(1:1000, 15I0053, affinity), MEX3D(1:1000, BE01081275, BIOSS), Signal detection was performed using an ECL chemiluminescence system (Epizyme).

Techniques:

Journal: Scientific Reports

Article Title: Comprehensive bioinformatics analysis of MEX3 family genes in hepatocellular carcinoma

doi: 10.1038/s41598-025-02057-0

Figure Lengend Snippet: The top 10 positively and top 10 negatively correlated genes for MEX3 family genes including MEX3A ( A ), MEX3B ( B ), MEX3C ( C ) and MEX3D ( D ). (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Membranes were blocked at room temperature with 5% non-fat milk for 2 h, then incubated overnight at 4 °C with primary antibodies: anti-MEX3A(1:800, CBG122A, CUSABIO) and anti-β-actin (1:2000, 20536-1-AP, Proteintech), MEX3B(1:1000, SA240318 × 09, ORIGEN), MEX3C(1:1000, 15I0053, affinity), MEX3D(1:1000, BE01081275, BIOSS), Signal detection was performed using an ECL chemiluminescence system (Epizyme).

Techniques: