FIGURE 9.

ASC with suppressed heme oxygenase 1 expression generate ASC-CM with reduced apoptosis-protective potency. (A) Left: ASC were treated continuously with 50 nM of non-specific (NS) or heme oxygenase 1 (HO-1)-specific siRNA for 48 h. Right: Treatment was applied for 24 h, followed by a 24 h incubation in siRNA-free media. Cell lysates were analyzed with antibodies to heme oxygenase 1 and β-actin (loading control). (B) ASC-CM was generated from ASC treated for 24 h with 50 nM non-specific RNA (NS RNA) or heme oxygenase 1-specific siRNA (siRNA), followed by siRNA removal and 24 h conditioning with media. HPAEC pre-incubated for 48 h with growth media or ASC-CM were stimulated with 2 ng/ml TNF. Cell lysates were analyzed with antibodies to cleaved and total caspase 3. Data from three independent experiments were pooled and presented as cleaved/total caspase 3 ratio mean ± SEM. ^∗Significance between TNF-stimulated groups assessed by t-test with Welch’s correction as indicated (p < 0.05).