Journal: Nature communications

Article Title: Suppression of TGF-β/SMAD signaling by an inner nuclear membrane phosphatase complex.

doi: 10.1038/s41467-025-58681-x

Figure Lengend Snippet: Fig. 5 | The CTDNEP1-NEP1R1 phosphatase and MAN1 suppress inappropriate SMAD signaling. A Localization of endogenous SMAD2 in HeLa parental cells or lacking the indicated genes analyzed by immunofluorescence. DNA was labeled with 4′,6- diamidino- 2- phenylindole (DAPI). Scale bar: 10 μM. B Quantification of nuclear accumulation of SMAD2 from imaging experiments, as shown in (A), n = 3 independent experiments, p-values were indicated in the graph. One-way ANOVA (multiple comparison) was performed, and data are presented as mean values ± SD. C Levels of endogenous pSMAD2 and SMAD2 in HeLa parental cells or lacking the indicated genes. Cell lysates were analyzed by SDS-PAGE followed by immuno- blotting with the indicated antibodies. GAPDH was used as a loading control. D Levels of endogenous pSMAD2 and SMAD2 in HeLa parental cells or CTDENP1 KO cells with indicated treatments. The 1D11 antibody against TGF-β cytokine was used at 30, 150, and 300 µg/mL; mouse IgG antibody was used at 300 µg/mL; the TGF-β receptor specific inhibitor SB431542 was used at 10 µM. Cell lysates were analyzed

Article Snippet: Inhibition of TGF-β with 1D11 neutralizing antibody Inhibition of TGF-β ligands was performed with the neutralizing antibody, 1D11 (BioX-Cell # #BE0083), and isotype-matched IgG1 monoclonal control antibody (BioX-Cell #BE0057) were used at 30, 150, and 300μg/mL, respectively, to treat HeLa parental cells or CTDENP1 KO cells with indicated time.

Techniques: Labeling, Imaging, Comparison, SDS Page, Control