Fig. 4
Nhlrc2FINCA/− mouse NPCs display a significant decrease in Nhlrc2 expression and in the amount of NHLRC2. a Immunoblotting and scatter plot of band intensities of NHLRC2 in four Nhlrc2+/+, four compound heterozygous Nhlrc2FINCA/−, three heterozygous Nhlrc2FINCA/+, and three heterozygous Nhlrc2+/− NPC whole-cell lysates. Nhlrc2FINCA/− (5%, p < 0.0001), Nhlrc2FINCA/+ (53%, p = 0.0062), and Nhlrc2+/− (37%, p = 0.0009) had significantly decreased NHLRC2 levels compared to wild-type NPCs. Nhlrc2FINCA/− also differed significantly from Nhlrc2FINCA/+ (p = 0.0007) and Nhlrc2+/− (p = 0.039) (Student’s t-test). Protein amounts are relative to one of the wild-type samples and GAPDH was used for normalization. b qPCR from four Nhlrc2FINCA/− and four Nhlrc2+/+ showed 50.2% mRNA expression (p = 0.0005) in the Nhlrc2FINCA/− NPCs when using primers designed over the intron between exons 4 and 5 of Nhlrc2, where the KO first allele construct resides and terminates the transcription. c qPCR, using a primer pair over the intron between exons 3 and 4 of Nhlrc2, from three Nhlrc2FINCA/− and three Nhlrc2+/+ NPCs, showed a significant decrease in Nhlrc2FINCA\− (70%, p = 0.0044) cells compared to Nhlrc2+\+ (Student’s t-test). Scatter plots show individual data points, group means, and SEM. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05