Figure 3.

Döhle‐like inclusion bodies localization by NMMHC‐IIA immunofluorescence or MGG staining. Light microscopy and immunofluorescence analyses of granulocytes in a healthy control (control), in patients (32 for immunofluorescence and 43 for light microscopy) with a pathogenic variant (pathogenic) and in three patients (28, 29, and 40) with a variant of uncertain significance (VUS). The analysis was performed by two independent centres: Panels I–V show results obtained by centre 1; Panels VI–X show results obtained by centre 2. Both centres used rabbit antihuman NMMHCIIA Ab followed by Alexa‐Fluor 488‐conjugated secondary antibody. Results between the two centers were highly comparable. The patient's sample in which a pathogenic variant was identified shows circular to oval shaped cytoplasmic punctuate spots, classified as type II inclusions (panels II and VII). Patients’ samples in which VUSs were identified show a speckled staining (panels III and VIII and panels V and X, respectively), and many small dots scattered throughout the cytoplasm (panels IV and IX) classified as type III inclusions. Panels XI–XV show May–Grünwald–Giemsa staining. Panels XII and XV show the presence of Döhle‐like bodies (arrowhead) in patients’ samples with a pathogenic variant (XII) and a VUS (XV). NMMHCIIA, nonmuscle myosin of class IIA; VUS, variant of uncertain significance