Image pro

To verify that all the nematodes belonged to the same species, we performed a detailed morphological investigation on a subset of the population (adults and juveniles). Several nematodes were mounted on slides for detailed morphological observation using the formalin–ethanol/glycerol technique^{54 (link),55} . Photos were captured on a Leica DM IRB microscope and a Zeiss AxioZoom microscope, each equipped with live camera (Image-Pro and Zen software, respectively).

YT cells were resuspended at 10⁶ cells mL⁻¹ in serum‐free RPMI‐1640 and adhered to glass slides for 30 min at 37°C, washed with PBS, containing 0.1% bovine serum albumin (BSA) (Sigma–Aldrich), 0.02% sodium azide and fixed with ice‐cold methanol for 5 min and washed in PBS before incubating for 1 h at room temperature in blocking buffer (PBS, 1% BSA). Samples were incubated with primary antibodies in PBS, 0.2% BSA for 1 h at room temperature, or overnight at 4°C and washed extensively in PBS, 0.2% BSA before adding secondary antibodies for 40 min at room temperature. Nuclei were stained with Hoechst 33342 (1:20,000) (Invitrogen, Thermofisher Scientific, Waltham, MA, USA) in PBS for 1–2 min before mounting with number 1.5 coverglass and mounting medium (Mowiol). Fixed images were examined at room temperature using either Zeiss LSM510 confocal microscope (Carl Zeiss, Inc., Oberkochen, Germany), or with laser‐scanning spinning‐disk confocal system (Andor Revolution) with lasers exciting at 405, 488, 543, and 633 nm using the 63x or 100x (Plan‐Apochromat, NA 1.40) oil immersion objective. Images were acquired using Image Pro (Zeiss, Inc.) or IQ software (Andor, Belfast, Northern Ireland).