Figure 2. Lysine suppresses PQ induced intracellular calcium
burden and EMT. (A) Co–IP analysis of STIM1 association with ORAI1 or
TRPC1 in A549 cells treated with 800 µM PQ for 0, 15, 30 min, together
w/wo 5 mM lysine as indicated. Images are representative of three
experiments. (B) NFAT luciferase expression in A549 cells
treated w/wo 800 µM PQ, together with 0-, 1-, 2.5-, or 5-fold lysine as
in the culture medium as indicated, for 24 hours. Mean ± s.d., ***P
< 0.001; One-Way ANOVA. n = 3. N.S., not significant.(C) Calcium imaging of A549 cells stimulated with 800 µM PQ,
followed by 2 mM CaCl2, and 100 µM, 200 µM, or 400 µM
lysine as indicated. Mean ± sem. (D-E) FACS analysis of
intracellular calcium levels by Fluo-3 staining in A549 cells (D) and
MLE 12 cells (E). Cells were treated w/wo PQ in a dose dependent manner
as indicated, together w/wo 5 mM lysine, for 24 hours. Mean ± s.d., ***P
< 0.001, *P < 0.05; unpaired two–tailed Student’st–test . n = 3. (F-H) WB analysis of the expression of
E-Cadherin and Vimentin, makers of EMT, in A549 cells (F), MLE-12 cells
(G), or A549 cells stably expressed STIM1 (H) treated with PQ in a dose
dependent manner, as indicated, together w/wo 5 mM lysine for 24 hours.(I-J) RT-PCR analysis to detect the expression ofE-Cadherin, or Vimentin in A549 cells (I) or MLE-12 cells (J)
treated with PQ in a dose dependent manner, as indicated, together w/wo
5 mM lysine for 18 hours.