Dfc420

A549 cells were seeded into six-well plates and transfected with siRNA of Gal-3 or treated by ICG-001. The cell monolayer was scraped in a straight line with a p200 pipette tip and incubated under serum-free conditions. Photographs of the scratch were taken under an invert microscope at 0, 12, 24, 36, and 48 h. Photographs at each time point were taken with Leica DFC420 camera. Gap width analysis was performed with scaleplate in microscope. Measurements were taken at multiple defined sites (>6) along the scratch. Each scratch was given an average of all measurements. Data are derived from three independent experiments.
Formula:
Migration distance ¼ ðGap width at 0 h ÀGap width at different time pointsÞ=Gap width at 0 h

Between July and December, 2013, 30 samples of P. arenatus with 20-63 (mean = 38.3) cm of length and with 400-3,600 (mean = 1,210) g of weight were obtained from fish markets in the cities of Niterói and Rio de Janeiro, state of Rio de Janeiro, Brazil.
The fish were transported in cool boxes containing ice to the Fish Inspection and Technology Laboratory of the Fluminense Federal University, where they were necropsied. The nematode larvae that were collected were fixed, clarified and preserved as described by Knoff & Gomes (2012) . The morphological analysis to identify H. deardorffoverstreetorum larvae was performed by Kuraiem et al. (2016) (link).
Small fragments of spleen, stomach, liver and mesentery tissues containing nodules and free parasites were extracted, fixed in 10% formaldehyde and processed for paraffin embedding (BEHMER et al., 1976) . Histological sections of 5 μm of thickness were mounted on microscope slides and stained with hematoxylin-eosin (HE) for subsequent analysis. Photomicrographs were obtained using a Leica DM1000 microscope coupled to a Leica DFC420 digital camera.
One slide showing a histopathological section through one of the parasites at the infection site was deposited in the Helminthological Collection of the Oswaldo Cruz Institute, Rio de Janeiro, RJ, Brazil, under the number 38207.

Intracellular reactive oxygen levels were quantified using the 2', 7'-dichlorodihydrofluorescein diacetate (H 2 DCFDA) molecular probes (Eugene, OR, USA) according to manufacturer instructions. Immediately after vitrification, ten embryos per treatment were washed twice in PBS, incubated for 60 minutes in 10 mM of H 2 DCFDA solution at room temperature, washed again in PBS and photographed using a Leica DM750 phase contrast microscope equipped with a Leica DFC420 camera. The wavelengths for excitation and emission were 492-495 nm and 517-527 nm, respectively. The percentage of surface area of the fluorescence-emitting embryo (affected by the formation of ROS) was measured using the software Image J (Schneider et al., 2012) (link). Culture medium (Leibowitz L-15) q.s. 100 mL