Fig. 4

TRIM27 is required for recruitment of JAK1 and STAT3 to the retromer. a Effects of VPS35 knockdown on the association of JAK1 with STAT3, gp130, TRIM27, or VPS35. HeLa cells (2 × 10⁷) were starved overnight followed by stimulation with IL-6 (50 ng/mL) for the indicated times. Coimmunoprecipitation and immunoblotting analysis were performed with the indicated antibodies. b Effects of TRIM27 knockdown on the association of JAK1 with VPS26, VPS35, or STAT3. HeLa cells (2 × 10⁷) were starved overnight followed by stimulation with IL-6 (50 ng/mL) for the indicated times. Coimmunoprecipitation and immunoblot analysis were performed with the indicated antibodies. c Effects of TRIM27 knockdown on the association of VPS26 with JAK1, STAT3, gp130, VPS35, or TRIM27. HeLa cells (2 × 10⁷) were starved overnight followed by stimulation with IL-6 (50 ng/mL) for the indicated times. Coimmunoprecipitation and immunoblotting analysis were performed with the indicated antibodies. d Effects of knockdown of the retromer components on IL-6-induced transcription of downstream genes. The control, VPS26-RNAi, VPS29-RNAi, and VPS35-RNAi stable HeLa cells (2 × 10⁵) were starved overnight and stimulated with IL-6 (20 ng/mL) for the indicated times before qPCR experiments. Data are representative of three experiments with similar results. Graphs show mean ± SD; n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, unpaired t test