Journal: bioRxiv

Article Title: Cell-nanoplastics association impacts cell proliferation and motility

doi: 10.64898/2026.04.03.716369

Figure Lengend Snippet: (A) Relative cell-associated NP fluorescence in MDCK II monolayers under treatment with different inhibitors. Cells were co-incubated with 20 µg/mL NPs and inhibitors for 6 hours. NP fluorescence intensity was normalized to vehicle control (DMSO). N = 3-6. (B) Dosedependent response of NP association under EIPA (NHE1 inhibitor) treatment. N = 6. (C) Relative NP release from MDCK II monolayers under various drug treatments. All samples were first incubated with 20 µg/mL NPs for 6 hours without drug treatment, followed by extensive washing and imaging to quantify the initial level of cell-associated NPs. Cells were then incubated in NP-free media containing inhibitors for 24 hours, after which NP fluorescence was measured again. The release fraction was calculated as the relative decrease in fluorescence intensity over this period. Conditions that compromised monolayer integrity due to cytotoxicity were excluded. N = 3-6. (D and E) Dose-dependent NP release fractions under treatment with CID-1067700 (Rab7 inhibitor; D) and LY294002 (PI3K inhibitor; E). N = 3-6. (F) Quantification of trans-epithelial NP transport sing a transwell system. MDCK II monolayers grown on 1 µm pore transwells were treated with 50 µg/mL NPs and inhibitors for 24 hours. NPs in the lower compartment were quantified using microfluidic devices. Empty transwells were used as baseline controls. N = 5. (G) Quantification of trans-epithelial NP transport in the presence of Dynasore or LY294002 using the assay in (F). N = 4-5. (H) Schematic illustration of pathways affecting NP-cell association. (A-G) Error bars represent standard deviation. (A and B) Paired t -test or RM one-way ANOVA was used. (C-G) Student’s t -test or one-way ANOVA was used.

Article Snippet: In selected experiments, cells were treated with the following pharmacological agents: vehicle control (0.25% DMSO; Invitrogen), Dynasore (Sigma), MyoVin-1 (Sigma), CID-1067700 (MilliporeSigma), EIPA (Tocris), syrosingopine (Cayman Chemical), Ouabain (Tocris), Valinomycin (Tocris), Gadolinium chloride (GdCl 3 ; Sigma), NSC668394 (MilliporeSigma), PF-562271 (Selleckchem), Y-27632 (Tocris), and LY-294002 hydrochloride (Sigma).

Techniques: Fluorescence, Incubation, Control, Imaging, Standard Deviation

Journal: bioRxiv

Article Title: Cell-nanoplastics association impacts cell proliferation and motility

doi: 10.64898/2026.04.03.716369

Figure Lengend Snippet: (A) Schematic overview of carboxylate-modified PS, PE, and PP NPs. (B and C) Quantification of cell-associated PS, PE, and PP NP density (B) after loading MDCK II monolayers with 20 µg/mL NPs for 24 hours, and NP release 24 hours after NP removal (C), following the method described in and . N = 4. (D and E) PS, PE, and PP NP association in MDCK II monolayers treated with EIPA (NHE1 inhibitor; D) or CID-1067700 (Rab7 inhibitor; E). N = 6. (F and G) NP association (F) and release fraction (G) under different viscosity conditions in MDCK II monolayers. (H) Relative cell number fold changes of NIH 3T3 cells cultured with 0, 2, 20, or 200 µg/mL NPs for 4 days. Cell number was quantified by DNA staining using Hoechst 33342. N = 3. (I and J) Migration speed and mean square displacement (MSD) of 3T3 fibroblasts on collagen-coated 2D substrates treated with 0, 20, 50, or 200 µg/mL PE (I) and PP (J) NPs for 6 hours. N = 3. (B-J) Error bars represent standard deviation. One-way ANOVA or Student’s t -test was used. RM one-way ANOVA was used in (D-F).

Article Snippet: In selected experiments, cells were treated with the following pharmacological agents: vehicle control (0.25% DMSO; Invitrogen), Dynasore (Sigma), MyoVin-1 (Sigma), CID-1067700 (MilliporeSigma), EIPA (Tocris), syrosingopine (Cayman Chemical), Ouabain (Tocris), Valinomycin (Tocris), Gadolinium chloride (GdCl 3 ; Sigma), NSC668394 (MilliporeSigma), PF-562271 (Selleckchem), Y-27632 (Tocris), and LY-294002 hydrochloride (Sigma).

Techniques: Modification, Viscosity, Cell Culture, Staining, Migration, Standard Deviation

Journal: mBio

Article Title: ATP1A1 enhances porcine reproductive and respiratory syndrome virus type 2 attachment and internalization

doi: 10.1128/mbio.03896-25

Figure Lengend Snippet: ATP1A1 facilitates PRRSV-2 entry via macropinocytosis and caveolae/raft-mediated endocytosis pathways. ( A–C ) The effect of CPZ (10 µM), EIPA (50 µM), and genistein (50 µM) on PRRSV-2 infection. Marc-145 cells with or without ATP1A1 knockdown were pre-treated with three inhibitors for 2 h, followed by PRRSV HuN4-F112 strain (MOI = 1) infection for another 12 h (with compounds present throughout). Then, the cell lysates were collected to detect viral RNA abundance via qRT-PCR. ( D–I ) Impact of ATP1A1 knockdown on endocytic marker uptake. Marc-145 cells with or without ATP1A1 knockdown were incubated with endocytic markers TFN-FITC (10 µg/mL), DTN-FITC (200 µg/mL), and CTB-AF488 (10 µg/mL) for 1 h. The cells were uninfected or infected with the PRRSV HuN4-F112 strain (MOI = 50). At 1 hpi, cells were collected, and the uptake rates were evaluated by flow cytometry. Representative images for TFN-FITC, DTN-FITC, and CTB-AF488 uptake are shown in panels D, E, and F with quantification provided in panels G, H, and I from three independent replicates. ( J and K ) Colocalization analysis of ATP1A1, PRRSV-2 particles, and DTN-positive macropinosomes or CTB-positive caveolae. Marc-145 cells were incubated with endocytic markers DTN-FITC (200 µg/mL, green) and CTB-AF488 (10 µg/mL, green) for 1 h. Then, the cells were uninfected or infected with the PRRSV HuN4-F112 strain (MOI = 50). At 1 hpi, cells were fixed and stained with rabbit anti-ATP1A1 pAb (red) and mouse anti-PRRSV N mAb (purple). Nuclei were labeled with DAPI (blue). Significant differences were indicated as follows: ns ( P > 0.05), * ( P < 0.05), ** ( P < 0.01), *** ( P < 0.001), and **** ( P < 0.0001).

Article Snippet: Various inhibitors, chlorpromazine hydrochloride (CPZ, HY-B0407A), genistein (GEN, HY-14596), ethylisopropylamiloride (EIPA, HY-101840), PP2 (HY-13805), Src inhibitor 1 (Src-l1, HY-101053), ouabain octahydrate (ouabain, HY-B0542), and rostafuroxin (PST2238, HY-12283) were purchased from MedChemexpress.

Techniques: Infection, Knockdown, Quantitative RT-PCR, Marker, Incubation, Flow Cytometry, Staining, Labeling