Constitutive knockdown of NLRP2 in H9 human embryonic stem cells. (A) NLRP2 gene expression in shLuc (control) and knockdown (KD) NLRP2 (128236, 130989, and 129801) H9 embryonic stem cells (ESCs), as determined by quantitative PCR. Values are mean ± standard deviation (SD) (n = 3). *P <0.01. (B) NLRP2 protein expression in shLuc and KD NLRP2 (128236 and 130989) H9 ESCs before (ESC) and during in vitro differentiation (days 10 (D10) and 20 (D20)), as determined by western blot. (C) Bright-field imaging and immunofluorescence staining of KD NLRP2 H9 ESCs. Scale bar = 50 μm. (D) Expression of pluripotent genes in shLuc and KD NLRP2 (128236 and 130989) H9 ESCs by quantitative PCR. Relative gene expression was first normalized to that of GAPDH, and then presented as the fold change relative to that in shLuc H9 ESCs. Values are mean ± SD (n = 3).