Journal: Journal of Translational Medicine

Article Title: Nordihydroguaiaretic acid inhibits bladder cancer metastasis through suppression of α1,3-mannosyltransferase expression and LRFN4 N-glycosylation

doi: 10.1186/s12967-025-06571-7

Figure Lengend Snippet: ALG3 promoted metastatic potential in bladder cancer through EMT and VM. A – D Transwell assays evaluating metastatic potential of T24/UMUC3 cells following ALG3 knockdown or ALG3 overexpression. Left: Representative images (Scale bars = 50 μm). Right: Quantified cell counts (10 random fields/well). E – H qRT-PCR analysis of EMT markers (E-cadherin, N-cadherin) showing ALG3-mediated transcriptional regulation. Data normalized to GAPDH and expressed as fold-change vs. controls. I Western blot confirmed ALG3-dependent protein-level changes in EMT markers after ALG3 siRNA or cDNA transfection. GAPDH served as the loading control. J Tube formation assays demonstrating ALG3’s role in promoting VM, as measured by total tube length, mesh and junction number. Scale bar = 50 μm. K – N qRT-PCR of VM markers (VE-cadherin, MMP14, MMP2 and MMP9) revealed ALG3-dependent regulation. O Western blot validated VM marker protein expression changes. GAPDH served as the loading control. * P <0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The slides were then incubated overnight at 4 °C in a humidified chamber with primary antibodies: ALG3 (DF14333, 1:100, Affinity Biosciences, Jiangsu, China), E-cadherin (A22850, 1:100, ABclonal Technology, Wuhan, China), N-cadherin (A19083, 1:100, ABclonal Technology, Wuhan, China), VE-cadherin (A25003, 1:200, ABclonal Technology, Wuhan, China), and MMP14 (A2549, 1:100, ABclonal Technology, Wuhan, China).

Techniques: Knockdown, Over Expression, Quantitative RT-PCR, Western Blot, Transfection, Control, Marker, Expressing

Journal: Journal of Translational Medicine

Article Title: Nordihydroguaiaretic acid inhibits bladder cancer metastasis through suppression of α1,3-mannosyltransferase expression and LRFN4 N-glycosylation

doi: 10.1186/s12967-025-06571-7

Figure Lengend Snippet: NDGA suppressed ALG3-drove metastasis in bladder cancer. A , B Transwell assays assessing cell migration and invasion capabilities in T24 and UMUC3 cells after ALG3 overexpression, or combined treatment with NDGA. Scale bars = 50 μm. C Tube formation assays evaluating vasculogenic mimicry formation ability of bladder cancer cells treated with ALG3 cDNA or combined with NDGA, assessing with total tube length, number of meshes and junctions. Scale bars = 50 μm. D Representative lung tissues from nude mice (n = 6/group) injected via tail vein with 1 × 10 6 oeNC or oeALG3 cells, indicating NDGA treatment (5 mg/kg/day, oral gavage) reduced lung colonization. E H&E staining visualizing metastatic foci in oeNC, oeALG3 and oeALG3 + NDGA groups, showing NDGA reduced metastasis incidence. Scale bars = 20μm. F , G Immunohistochemical staining of EMT markers (E-cadherin, N-cadherin) and VM markers (VE-cadherin, MMP14) in tumor tissues collected from nude mice model, corroborating anti-metastatic mechanism. Scale bars = 20 μm. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The slides were then incubated overnight at 4 °C in a humidified chamber with primary antibodies: ALG3 (DF14333, 1:100, Affinity Biosciences, Jiangsu, China), E-cadherin (A22850, 1:100, ABclonal Technology, Wuhan, China), N-cadherin (A19083, 1:100, ABclonal Technology, Wuhan, China), VE-cadherin (A25003, 1:200, ABclonal Technology, Wuhan, China), and MMP14 (A2549, 1:100, ABclonal Technology, Wuhan, China).

Techniques: Migration, Over Expression, Injection, Staining, Immunohistochemical staining