Table_1_Parallel phonological processing of Chinese characters revealed by flankers tasks.XLSX

An important and extensively researched question in the field of reading is whether readers can process multiple words in parallel. An unresolved issue regarding this question is whether the phonological information from foveal and parafoveal words can be processed in parallel, i.e., parallel phonological processing. The present study aims to investigate whether there is parallel phonological processing of Chinese characters. The original and the revised flankers tasks were applied. In both tasks, a foveal target character was presented in isolation in the no-flanker condition, flanked on both sides by a parafoveal homophone in the homophone-flanker condition, and by a non-homophonic character in the unrelated-flanker condition. Participants were instructed to fixate on the target characters and press two keys to indicate whether they knew the target characters (lexical vs. non-lexical). In the original flankers task, the stimuli were presented for 150 ms without a post-mask. In the revised flankers task, we set the stimulus exposure time (duration of the stimuli plus the blank interval between the stimuli and the post-mask) to each participant’s lexical decision threshold to prevent participants from processing the target and flanker characters serially. In both tasks, reaction times to the lexical targets were significantly shorter in the homophone-flanker condition than in the unrelated-flanker condition, suggesting parallel phonological processing of Chinese characters. In the revised flankers task, accuracy rates to the lexical targets were significantly lower in the unrelated-flanker condition compared to the homophone-flanker condition, further supporting parallel phonological processing of Chinese characters. Moreover, reaction times to the lexical targets were the shortest in the no-flanker condition in both tasks, reflecting the attention distribution over both the target and flanker characters. The findings of this study provide valuable insights into the parallel processing mechanisms involved in reading.</p

NRP1 may play a role in the regulation of proliferation and drug resistance in OSCC cells.

<p>(<b>A</b>) Images of colonies of CAL27-P/N, HN4-P/N, and HN6-P/N cells stained with crystal violet. (<b>B</b>) Flow cytometric analysis of apoptosis in CAL27-P/N, HN4-P/N, and HN6-P/N cells treated with different concentrations of cisplatin for 24 h. (<b>C</b>) Quantification of cell colonies. Each data point represents the mean ± SD of data from three independent trials. *P<0.05; **P<0.01. (<b>D</b>) Statistical analysis of the rate of cell apoptosis in CAL27-P/N, HN4-P/N, and HN6-P/N cells. Each bar represents the mean ± SD of data from three independent trials. *P<0.05; **P<0.01.</p

High expression of NRP1 correlates to lymph node metastasis and poor prognosis in OSCC patients.

<p>(<b>A</b>) The expression of NRP1 in normal oral epithelium (a), a tissue sample from a patient without lymph node metastasis (b), and a tissue sample from a patient with lymph node metastasis (c) (magnification, ×200). (<b>B</b>) Kaplan-Meier overall survival curves for 60 patients with OSCC, according to NRP1 expression. *P<0.05.</p

Association between OSCC clinical-pathological parameters and NRP1 expression.

<p>N, negative; L, low expression; H, high expression; N0, no nodal metastasis; N+, nodal metastasis.</p><p>*P<0.05.</p

Inhibition of the NF-κB signaling pathway results in a reversion of NRP1-mediated EMT.

<p>After treatment with PDTC for 24(<b>A, B</b>) western blotting and (<b>C</b>) Real time RT-PCR was used to detect the expression of EMT-related proteins in CAL27-P/N, HN4-P/N, and HN6-P/N cells. β-actin and histone H1 were employed as controls. Each data point represents the mean ± SD of three independent trials. *P<0.05; **P<0.01.</p

The examination of the biological properties of OSCC cells transfected with an empty vector or a vector encoding NRP1.

<p>(<b>A</b>) The differences in the migration ability between the CAL27-P, HN4-P, and HN6-P and CAL27-N, HN4-N, and HN6-N cells, respectively, were measured using the scratch assay. The CAL27-N, HN4-N, and HN6-N cells grew into the wounded area after 48 h, while CAL27-P, HN4-P, and HN6-P cells did not. The images were captured at ×40 magnification; Scale bar, 100 µm. (<b>B</b>) A transwell assay was employed to analyze the cell invasion ability. CAL27-N, HN4-N, and HN6-N cells had significantly higher invasion ability compared with CAL27-P, HN4-P, and HN6-P cells. The images were captured at ×200 magnification; Scale bar, 100 µm. Quantitative analysis of the data from the (<b>C</b>) scratch assay after 48 h and (<b>D</b>) transwell invasion assays after 24 h in five randomly-selected fields. The data shown are the mean ± SD. *P<0.05; **P<0.01.</p

Expression of NRP1 in normal oral epithelium and OSCC.

<p>N, negative; L, low expression; H, high expression.</p><p>*P<0.05.</p

Inhibition of NF-κB signaling pathway alters the biological properties of NRP1-expressing OSCC cells.

<p>(<b>A</b>) Scratch assays were performed to examine the migration ability in CAL27-P/N, HN4-P/N, and HN6-P/N cells, treated with vehicle control or PDTC for 24 h. Images of the wound area were captured at ×40 magnification; Scale bar, 100 µm. (<b>B</b>) Transwell assays with a reconstituted basement membrane were performed to examine the invasion ability in CAL27-P/N, HN4-P/N, and HN6-P/N cells, treated with vehicle control or PDTC for 24 h. The images were captured at ×200 magnification; Scale bar, 100 µm. Quantitative analysis of data from the (<b>C</b>) scratch assays after 48 h and (<b>D</b>) cell invasion assays after 24 h in five randomly-selected fields. The data shown are the mean ± SD of three independent trials. *P<0.05; **P<0.01.</p

NRP1-mediated EMT occurs through activation of the NF-κB signaling pathway.

<p>(<b>A</b>) Western blotting was performed to assess the expression level of NF-κB pathway-related proteins in cytoplasmic and nuclear extracts from CAL27-P/N, HN4-P/N, and HN6-P/N cells. β-actin and histone H1 were employed as the positive controls for cytoplasmic and nuclear proteins, respectively. (<b>B</b>) Semi-quantitative analysis of changes in protein expression as determined by densitometric scanning of the immunoreactive bands. Each data point represents the mean ± SD of three independent trials. *P<0.05; **P<0.01.</p

NRP1 induces the EMT process in OSCC cell lines.

<p>(<b>A</b>) CAL27, HN4, and HN6 cells transfected with the vector control (CAL27-P, HN4-P, and HN6-P) had a typical pebble-like epithelial morphology, while cells transfected with the NRP1-expressing plasmid (CAL27-N, HN4-N, and HN6-N) had a more elongated, pebble-like morphology. The images were captured using an inverted microscope (Olympus) fitted with a camera. The images were captured at ×40 magnification; Scale bar, 100 µm. (<b>B, C</b>) Western blotting and (<b>D</b>) Real time RT-PCR analyses were used to assess the expression of epithelial (E-cadherin and β-catenin) and mesenchymal (N-cadherin and vimentin) markers in OSCC cells transfected with the vector control or a vector encoding NRP1. β-actin and GAPDH were employed as controls in the western blotting and RT-PCR analyses, respectively. Each data point represents the mean ± SD of three independent repetitions of the experiment. *P<0.05; **P<0.01.</p