prmt6  (BPS Bioscience)

Journal: Journal of experimental & clinical cancer research : CR

Article Title: RBM10 deficiency promotes brain metastasis by modulating sphingolipid metabolism in a BBB model of EGFR mutant lung adenocarcinoma.

doi: 10.1186/s13046-025-03347-1

Figure Lengend Snippet: Fig. 4 RBM10 regulates the biosynthesis of sphingolipid metabolites. (A) KEGG pathway analysis of differentially expressed genes. (B) Volcano plot for dif ferentially expressed genes. (C) Molecular docking simulation of RBM10 with GALC. (D) Metabolite heat map clustering results. (E) Volcano plot showing fold changes of metabolites. (F) Heat map analysis revealing the correlation between SPH and Cer

Article Snippet: Immunohistochemistry (IHC) was conducted following the method previously described. (21) For the primary antibody incubation, RBM10 (HPA034972, Sigma,1:300) and GALC (11991-AP, Proteintech,1:150) were employed for IHC.

Techniques:

Journal: Journal of experimental & clinical cancer research : CR

Article Title: RBM10 deficiency promotes brain metastasis by modulating sphingolipid metabolism in a BBB model of EGFR mutant lung adenocarcinoma.

doi: 10.1186/s13046-025-03347-1

Figure Lengend Snippet: Fig. 5 RBM10 regulates GALC, a key enzyme in the sphingolipid metabolic pathway, through alternative splicing. (A) WB was used to detect RBM10 and GALC expression after treatment with madrasin. (B) New transcripts were detected by EU incorporation assay. (C) T7-Ftz reporter minigene assay. (D) Nanopore sequencing analysis showed the numbers of altered splicing events in each AS category. (E-F) Exon 6 skipping in GALC was identified through a comprehensive analysis. (G) The RBM10 binding motif was obtained from MACS2. (H) Bubble map annotated by KEGG pathway analysis. *P < 0.05, **P < 0.01

Article Snippet: Immunohistochemistry (IHC) was conducted following the method previously described. (21) For the primary antibody incubation, RBM10 (HPA034972, Sigma,1:300) and GALC (11991-AP, Proteintech,1:150) were employed for IHC.

Techniques: Alternative Splicing, Expressing, Mini Gene Assay, Nanopore Sequencing, Binding Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: RBM10 deficiency promotes brain metastasis by modulating sphingolipid metabolism in a BBB model of EGFR mutant lung adenocarcinoma.

doi: 10.1186/s13046-025-03347-1

Figure Lengend Snippet: Fig. 6 RBM10 inhibits GALC-L and promotes GALC-S to exert tumor-suppressor function. (A, C) Minigene assays showing the splicing status. (B, D) Quantification of exon 6 inclusion levels is shown. PSI: percent-spliced-in. (E) Expression of GALC proteins after transfection with siGALC-L and siGALC-S were examined by WB. (F-G) Trans-BBB migration assay was used to test the transfer capacity under siGALC-L and siGALC-S conditions. (H-I) The S1P con centration in tumor cells was quantified using the enzyme-linked immunosorbent assay (ELISA). Scale bar: 10x:50 μm. *P < 0.05, **P < 0.01, **** P < 0.0001

Article Snippet: Immunohistochemistry (IHC) was conducted following the method previously described. (21) For the primary antibody incubation, RBM10 (HPA034972, Sigma,1:300) and GALC (11991-AP, Proteintech,1:150) were employed for IHC.

Techniques: Expressing, Transfection, Migration, Enzyme-linked Immunosorbent Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: RBM10 deficiency promotes brain metastasis by modulating sphingolipid metabolism in a BBB model of EGFR mutant lung adenocarcinoma.

doi: 10.1186/s13046-025-03347-1

Figure Lengend Snippet: Fig. 7 RBM10 deficiency promotes GALC migration to the nucleus. (A) ChIP analysis of Pol II-pSer2 binding to the GALC promoter during RBM10 deple tion (Pol II-pSer2: RNA polymerase II CTD repeat YSPTSPS antibody). (B) WB assay of nuclear and cytoplasmic cellular fractions used to detect the nuclear translocation of GALC. β-actin was used as the cytoplasmic control, and Lamin B1 as the nuclear control. (C) Expression of GALC (green) in PC9BrM3 and 3255BrM3 cells when RBM10 was down-regulated. Nuclei counterstained with DAPI (blue). (D-E) In vivo co-localization of the RBM10 and GALC. (F) RIP assay to verify the binding between RBM10 and GALC. (G) RNA pull-down assay showing that RBM10 binds to GALC exon 6 in vitro. Scale bar: 20x:40 μm, 40x:20 μm, 60x:10 μm. **P < 0.01, ***/****P < 0.001

Article Snippet: Immunohistochemistry (IHC) was conducted following the method previously described. (21) For the primary antibody incubation, RBM10 (HPA034972, Sigma,1:300) and GALC (11991-AP, Proteintech,1:150) were employed for IHC.

Techniques: Migration, Binding Assay, Translocation Assay, Control, Expressing, In Vivo, Pull Down Assay, In Vitro