Fig. 5

POSTN directly binds to membrane receptor NOTCH1 to promote the activation of hepatic stellate cells. A Schematic diagram of POSTN and NOTCH1 truncations; B-C Co-IP was used to analyze the interaction between POSTN and full-length NOTCH1 (B) or NOTCH1-NECD domain (C) in 293T cells; D Pull down FLAG to analyze the interaction between NOTCH1-NECD domain and different POSTN truncations in 293T cells; E Pull down HA to analyze the interaction between NOTCH1-NECD domain and different POSTN truncations in 293T cells; F-G The co-localization of POSTN full-length or truncations and NOTCH1-NECD domain in 293T cells detected by Immunofluorescence analysis (F). Quantification of the Pearson value was shown (G). Scale bar = 10 μm; H-I. I The co-localization of exogenous POSTN and NOTCH1-NECD domains in LX-2 cells treated with CM from POSTN-FLAG-overexpression H446 cells detected by Immunofluorescence analysis (H). Quantification of the fluorescence intensity was shown (I). Scale bar = 10 μm; J The indicated proteins level in LX-2 cells treated with SCLC CM and/or siNOTCH as indicated for 48 h; K The indicated proteins level in LX-2 cells treated with SCLC CM and/or DAPT (20 μM) as indicated for 48 h. *P < 0.05, **P ≤ 0.01, ***P < 0.001, ****P ≤ 0.0001. n = 3. Student's t-test. All data were shown as means ± sem