Figures & data
Fig. 1. Lewis lung carcinoma (LLC)-derived conditioned medium (CM) decreased Semaphorin-3a (Sema3a) expression in MC3T3-E1 cells under low-serum culture conditions.
Notes: LLC cells were cultured in DMEM containing (A) 10% or (B) 1% FBS for 24 h, and CM was harvested. MC3T3-E1 cells were treated with LLC-CM or cell free-CM (DMEM) for 24 h, total RNA was extracted, and then reverse-transcribed to cDNA. Sema3a level was analyzed using qRT-PCR. Values are normalized to Actb level. n = 3, ***p<0.001 (Student’s t-test). DMEM, Dulbecco’s modified Eagle’s medium; FBS, fetal bovine serum; qRT-PCR, quantitative real-time reverse transcription-polymerase chain reaction.
Fig. 2. Rapamycin restored decrease in Semaphorin-3a (Sema3a) expression in MC3T3-E1 cells induced by Lewis lung carcinoma conditioned-medium (LLC-CM) treatment.
Notes: MC3T3-E1 cells were treated with LLC-CM, 1 nM rapamycin, or LLC-CM/1 nM rapamycin in 1% FBS/DMEM for 24 h. (A) Western blots of phosphorylated (p) p70S6 K (T389), p70S6 K, and actin in MC3T3-E1 cells treated with DMEM or LLC-CM. (B) Quantification of pp70S6 K/p70S6 K ratio. n = 3, ***p<0.001 (Student’s t-test). (C) Total RNA was reverse-transcribed to cDNA. Sema3a level was analyzed using qRT-PCR. Values are normalized to Actb level; *p < 0.05 (Student’s t-test), n = 3. FBS, fetal bovine serum; DMEM, Dulbecco’s modified Eagle’s medium; qRT-PCR, quantitative real-time reverse transcription-polymerase chain reaction.
