Research Paper Volume 12, Issue 16 pp 16326—16340

Figure 4. PTEN blocks TNF-α-induced Ca²⁺ accumulation and CREB activation by downregulating CD38. ASM cells infected with Ad-CD38 shRNA were incubated for 48 h and stimulated with of 20 ng × ml^-1 TNF-α for 24 h, and (A) intracellular Ca²⁺ concentrations were measured with Fluo-3 AM fluorescent probes, and (B, C) the protein expression patterns of total CREB and phosphorylated CREB (pCREB) were determined via Western blot (n = 3, **p < 0.01 ***p < 0.001 and ****p < 0.0001). ASM cells were infected with Ad-PTEN or/and Ad-CD38, incubated for 48 h, and stimulated using of 20 ng × ml^-1 TNF-α for 24 h. (D) Intracellular Ca²⁺ concentrations were assessed using Fluo-3 AM fluorescent probes, and (E, F) protein expression levels of total CREB and phosphorylated CREB (pCREB) were determined via Western blot (n = 3, **p < 0.01 ***p < 0.001 and ****p < 0.0001).