Fig. 4.

Nedd4L impairs LPA-LPA1 signaling. a. MLE12 cells were transfected with HA-nedd4l plasmids (0–2 μg) for 48 h, and then cells were treated with LPA (5 μM) for 10 min. Cell lysates were analyzed by immunoblotting with p-p38 MAPK, p38 MAPK, p-Erk1&2, Erk1&2, HA, and β-actin antibodies. b. MLE12 cells were transfected with Cont shRNA or Nedd4l shRNA for 72 h, and then cells were treated with LPA (5 μM) for 0–60 min. Cell lysates were analyzed by immunoblotting with p-p38 MAPK, p38 MAPK, p-Erk1&2, Erk1&2, Nedd4L, and β-actin antibodies. Representative immunoblots were from at least three independent times. c. HBEpCs were transfected with HA-NEDD4L plasmid for 48 h, and then cells were treated with LPA (1 μM) for 3 h. Cells were collected and IL-8 mRNA levels were analyzed by RT-realtime PCR.