Figure 8.

AAD-induced FAM134B2 expression is calcium dependent. (A–C) FAM134B2, MEF2D and NR4A1 mRNA expression in HeLa cells treated with HisOH in the presence of BAPTA. HeLa cells were pretreated with 20 μM BAPTA-AM for 2 h and then treated with 2 mM HisOH for 6 h. (D, E) FAM134B2 and DDIT3 mRNA expression in HeLa cells treated with thapsigargin (TG), tunicamycin (TUN) and ionomycin (ION). HeLa cells were treated with 0.5 mM thapsigargin, tunicamycin and ionomycin for 6 h. (F) Measurement of cytosolic Ca²⁺ with Fluo-4 DirectTM. HeLa cells in a 96-well microplate were incubated with complete media and 2x Fluo-4 DirectTM reagent for 1 h, and then fluorescence was measured with a microplate reader at every 1 min. Cells were treated with ddH₂O (Control) or 2 mM HisOH for 7 min after the start of the measurement. Data was expressed as the mean of fold value against average intensity at pretreatment (values from 0 to 4 min) ±SEM. (n = 4), G) FAM134B2, H) MEF2D, I) NR4A1 and J) DDIT3 expression in HeLa cells treated with HisOH in the presence of PKC and CAMK inhibitor. HeLa cells were pretreated with 10 μM Go6983 or KN-93 for 2 h and then treated with 2 mM HisOH for 6 h. *P < 0.05 and ***P < 0.001.