Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A mRNA level of RTN3 in thyroid cancer and normal tissues from TCGA database. B Relationship between RTN3 expression and pathologic T stage in thyroid cancer patients from TCGA database. C Kaplan–Meier survival analysis of progress free survival in patients with low or high RTN3 expression from TCGA-THCA cohort. D Association of RTN3 and gene set of thyroid cancer dedifferentiation (FOXE1, NKX2-1, NIS, PAX8, TG, TPO and TSHR). E Representative IHC staining images of RTN3 (left) and relative expression levels (right) in 45 pairs of thyroid cancer and adjacent normal tissues (scale bar, 100 μm). F , G qRT-PCR ( F ) and WB ( G ) detected mRNA and protein expression levels of RTN3 in normal thyroid and thyroid cancer cell lines. H , Validation of RTN3 knockout by WB. I –K Cell proliferation ability after RTN3 knockout determined by colony-formation assays ( I ) , CCK-8 assays ( J ) and EdU assays ( K ) with representative images in the left (scale bar, 40 μm) and the quantification of EdU positive cell ratios in the right, n = 3. L , M Cell migration ability after RTN3 knockout determined by transwell assays ( L ) (scale bar, 40 μm) and wound healing assays ( M ) (scale bar, 100 μm), with representative images in the left and the quantification graphs in the right, n = 3. *: P < 0.05, **: P < 0.01, ***: P < 0.001, ****: P < 0.0001.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR, Biomarker Discovery, Knock-Out, CCK-8 Assay, Migration

Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A , B Validation of the overexpression efficiency of RTN3 plasmids by qRT-PCR ( A ) and WB ( B ) in KTC-1 and K1 cells. C –E Cell proliferation ability of RTN3 overexpression cells determined by CCK-8 assays ( C ), EdU assays ( D ) with representative images in the left (scale bar, 40 μm) and the quantification of EdU positive cell ratios in the right, and colony-formation assays ( E ), n = 3. F , G Cell migration ability of RTN3 overexpression cells determined by transwell assays ( F ) (scale bar, 40 μm) and wound healing assays ( G ) (scale bar, 100 μm), with representative images in the left and the quantification graphs in the right, n = 3. *: P < 0.05, **: P < 0.01, ***: P < 0.001, ****: P < 0.0001, ns: P ≥ 0.05.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Biomarker Discovery, Over Expression, Quantitative RT-PCR, CCK-8 Assay, Migration

Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A Mass spectrometry assay depicted the DHCR7 peptides pulled down by anti-RTN3 antibody. B WB analysis of IP assays using the indicated antibodies conducted in 293 T cells transiently transfected with the indicated plasmids. C, D WB analysis of IP assays using RTN3 antibody conducted in KTC-1 cells ( C ) and K1 cells ( D ). E Schematic representation of truncated RTN3. F WB analysis of IP assays using the anti-FLAG antibody conducted in 293 T cells transiently transfected with the truncated RTN3 plasmids.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Mass Spectrometry, Transfection

Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A WB analysis of DHCR7 expression levels after knockout, knockdown and overexpression of RTN3 in KTC-1 (left) and K1 (right) cells. B IHC analysis of the correlation between RTN3 and DHCR7 protein levels of thyroid cancer tissues. Upper: Representative staining results of low and high expression of RTN3 and DHCR7; scale bar, 100 μm. Lower: Correlation analysis showed negative relationship between RTN3 and DHCR7 proteins. C mRNA levels of DHCR7 in RTN3 knockout, knockdown and overexpression KTC-1 (left) and K1 (right) cells. ns: P ≥ 0.05. D WB analysis of DHCR7 and RTN3 levels after CHX treatment for indicated time in RTN3 knockout (upper) or overexpression (lower) cells. E WB analysis of the levels of RTN3 and DHCR7 after vehicle control or MG132 treatment (20 μg/mL, 8 h) in RTN3 knockout cells. F Cells with RTN3 knockout or overexpression were transfected with Ub-HA plasmids and treated with MG132 (20 μg/mL, 8 h) 64 h after transfection. IP and WB assays using indicated antibodies were used to analyze the ubiquitination of DHCR7.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Expressing, Knock-Out, Knockdown, Over Expression, Staining, Control, Transfection, Ubiquitin Proteomics

Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A Cholesterol concentrations in KTC-1 (left) and K1 (right) cells with RTN3 knockout, knockdown and overexpression, n = 3. B The effects of cholesterol and its inhibitor Simvastatin on the activation of EGFR/ERK pathway indicated by p-EGFR and p-ERK levels detected by WB assay in KTC-1 (left) and K1 (right) cells. C WB analysis of p-EGFR and p-ERK levels after RTN3 knockout, knockdown and overexpression in KTC-1 (left) and K1 (right) cells. D WB analysis of the effects of Simvastatin treatment on p-EGFR and p-ERK levels in RTN3 knockout cells. E –G Cell proliferation ability determined by CCK-8 assays ( E ) and EdU assays ( F ) with representative images in the left (scale bar, 40 μm) and the quantification of EdU positive cell ratios in the right, and colony-formation assays ( G ), n = 3. H , I Cell migration ability determined by transwell assays ( H ) (scale bar, 40 μm) and wound healing assays ( I ) (scale bar, 100 μm), with representative images in the left and the quantification graphs in the right, n = 3. *: P < 0.05, ***: P < 0.001, ****: P < 0.0001, ns: P ≥ 0.05.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Knock-Out, Knockdown, Over Expression, Activation Assay, CCK-8 Assay, Migration

Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A Drug sensitivity of Mirdametinib, Selumetinib, and Trametinib of KTC-1 cells and their IC50 after RTN3 knockout, n = 3. B Proliferation curve of KTC-1 cells with treatment of Mirdametinib at 20 nM. C The effects of Mirdametinib on the protein levels of p-ERK after RTN3 knockout. D The scheme of in vivo xenograft assay and Mirdametinib treatment. E , F Tumor volume ( E ) and growth curve (measured every 2 days) ( F ). G Representative images (left; scale bar, 100 μm) and quantification (right) of IHC staining of RTN3 and Ki-67. n = 5.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Knock-Out, In Vivo, Xenograft Assay, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: Declined RTN3 stabilizes DHCR7 to induce cholesterol-dependent tumor progression and MEK inhibitors insensitivity in thyroid cancer

doi: 10.1038/s41419-026-08538-y

Figure Lengend Snippet: A , B siDHCR7 ( A ) and Simvastatin ( B ) rescued insensitivity to Mirdametinib caused by RTN3 KO. n = 3. C , D Tumor volume ( C ) and growth curve (measured every 2 days) ( D ) of xenograft tumors. E , F Representative images ( E ) (scale bar, 100 μm) and quantification ( F ) of IHC staining of RTN3 and Ki-67. n = 5. *: P < 0.05, ****: P < 0.0001, ns: P ≥ 0.05.

Article Snippet: The following antibodies were used: DDDDK-Tag (AE005, Abclonal), His-Tag (10001-0-AP, Proteintech), RTN3 (68215-1-Ig, Proteintech), Rabbit IgG (30000-0-AP, Proteintech), Mouse IgG (B900620, Proteintech).

Techniques: Immunohistochemistry