Identification of SRPRA and SRP19 novel gene variants. (A) Chest computed tomography scan of index patient II-3 (family A). (B) Transelectron microscopy sections of neutrophils from index patient A.II-3 compared with unaffected family members (brother A.II-2, father A.I-2) and healthy donors. Non-adherent Neutrophils have a size of ca. 9 μm. (C) Quantification of neutrophil granule content from healthy donor, A.I-2, A.II-2, and A.II-3. Group differences with P < .0001. Single-group differences via Student t test. ∗P < .01, ∗∗P < .001, and ∗∗∗∗P < .00001. (D) Pedigree of family A and Sanger sequencing chromatograms of wild-type (WT) and the SRPRA mutation site. Ribbon representation of the 3-dimensional structure of the human SRPRA WT (E) and mutated SRPRA (Q464E) (F). (G) Pedigree of family B and Sanger sequencing chromatograms of the WT and the SRP19 mutation site. (H) Reverse transcription polymerase chain reaction (RT-PCR) results documenting WT and mutated (mut) SRP19_2 to 4 minigene transcripts in HeLa cells. F1-R1 and F1-R2 are the primer pairs used to amplify the indicated SRP19 exons. The primer pair F1-R2 amplifies SRP19 exon 2 to 4 as indicated with a red arrow (281 base pair [bp]). The SRP19 variant results in a PCR product of only 209 base pair (blue arrow). The primer pair F1-R1 amplifies parts of the SRP19 exon 2 and is used as a transfection efficiency control. (I) Immunoblot analysis of EBV-LCL lysates from healthy donor, patient (Pat.) B.V-1, and a heterozygous family (fam.) member (B.IV-6). Experiment performed in triplicate.