Jem 1400 80 kv tem

TEM: the ferritin samples (10 μL, 0.1 mg mL^-1) were embedded in a Plasma Cleaner HPDC32G-treated copper grid and stained with 1% uranyl acetate for 1 min, then imaged with a JEM-1400 80 kV TEM (JEOL, Japan). DLS: the average diameter of the ferritin samples (20 μL, 0.1 mg mL^-1) was analyzed by the DynaPro Titan with a temperature-controlled micro-sampler of 25 °C (Wyatt Technology, U.S.A.). Inductively coupled plasma mass spectrometry (ICP-MS): the element contents of ferritin samples from different species were analyzed by the Agilent ICPMS7800.

The tomograms were acquired from serial sections of resin‐embedded (Spurr) mosquito midguts. Each section (100 and 300 nm thickness for WT and α1‐tubulin(‐) parasites, respectively) was inspected for suitable objects and mapped using a JEOL JEM‐1400 80 kV TEM. The tilt series were performed on a FEI Tecnai F30 300 kV TEM with the Gatan OneView sensor (Gatan Inc, Pleasanton, CA, USA) installed and controlled by SerialEM (Mastronarde, 2005). Each series ranged from ± 60–70° with images at 2° increments at 9,600× magnification. The tomogram volumetric reconstruction for each individual section was performed using the IMOD 4.9 software package (Kremer et al, 1996). Every image in the tilt series was aligned and tracked via patch tracking, and the volume was reconstructed using weighted‐back projection. The 3D reconstructions of the sections were flattened and trimmed before combining them into a single volume. For visual representation, the objects of interest in each tomogram were manually segmented in 3dmod (IMOD). The animations (Movies EV1 and EV2) were created by exporting frames in 3dmod and combining them using FFmpeg (FFmpeg Developers, version: be1d324).