Fig. 3

HMGB1-mediated autophagy regulated NIS expression and iodide uptake (a). FTC-133/TPC-1 cells were pretreated with 3-MA (10 mM) treatment for 1 h and then starved by HBSS for 3 h. LC3-I/II and NIS levels were assayed by Western blot; (b&c) FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and then starved by HBSS for 3 h. LC3-I/II and NIS levels were assayed by Western blot. And NIS mRNA was assayed by qRT-PCR (n = 3, *P > 0.01, **P > 0.01); (d) Dynamic uptaking: FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and starved by HBSS for 3 h. Indicated cells were cultured with 175 KBq ¹³¹I for 5, l0, 20, 30, 60, 90 and 120 min. The uptake of ¹³¹I in indicated cells was detected by a gamma counter; (e) Radionuclide uptaking: FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA in the presence or absence of 3-MA (10 mM) treatment for 1 h and then starved by HBSS for 3 h. After 1-h incubating with ¹³¹I, the uptake of ¹³¹I in indicated cells was detected by a gamma counter (n = 3, *P < 0.01, **P > 0.01); (f) FTC-133/TPC-1 cells transfected with HMGB1 shRNA and control shRNA were starved by HBSS for 3 h and then treated with rapamycin (1 μM) for 12 h. After 1-h incubating with ¹³¹I, the uptake of ¹³¹I in indicated cells was detected by a gamma counter. Rap, rapamycin. (n = 3, *P < 0.01, **P > 0.01)