nih3t3 mouse embryonic fibroblasts  (ATCC)

Journal: ACS applied materials & interfaces

Article Title: Cellular Uptake of Nanoparticles is Regulated by Integrin-Based Adhesion to the Extracellular Matrix

doi: 10.1021/acsami.5c18030

Figure Lengend Snippet: Characterization of cell spread area and focal adhesion formation on different coatings via immunostaining of cells after 24h incubation. (A) Cell spread area for NIH3T3 cells with (B) quantification of average focal adhesion size (n=39–40 cells per group from 3 independent experiments). (C) Cell spread area for RAW264.7 cells (n=45 cells per group from 3 independent experiments) and (D) ASCs with (E) quantification of average focal adhesion size (FAAS) (n=45 cells per group from 3 independent experiments). ( F ) Representative fluorescence images of focal adhesion formation via paxillin staining of NIH3T3, RAW264.7, and ASCs cells on different coating types. ( G ) Representative fluorescence images of cytoskeletal arrangement via phalloidin staining of NIH3T3, RAW264.7, and ASCs cells on different coating types. Violin plots indicate mean, median, quartiles and distribution (width). Bars indicate mean ± SEM. Cell area analyzed by One-way ANOVA with Tukey’s multiple comparison test. Symbols for statistical comparisons between indicated groups: * p ≤ 0.05; **p ≤ 0.01; or ## p<0.01 vs. all other groups. (scale bars = 40μm).

Article Snippet: Three different cell types, NIH3T3 fibroblasts (ATCC, Manassas, VA, USA, Cat. No. CRL-1658), RAW264.7 macrophages (ATCC, Manassas, VA, USA, Cat. No. TIB-71), and primary rat adipose-derived stem cells (ASCs), isolated from female 8–14 week-old Lewis rats (Charles River Laboratories, Charleston, SC, USA) were used.

Techniques: Immunostaining, Incubation, Fluorescence, Staining, Comparison

Journal: ACS applied materials & interfaces

Article Title: Cellular Uptake of Nanoparticles is Regulated by Integrin-Based Adhesion to the Extracellular Matrix

doi: 10.1021/acsami.5c18030

Figure Lengend Snippet: Images of PLGA nanoparticle (Green) uptake and quantification for NIH3T3 cells( A, B ), RAW264.7 cells( C, D ), and ASCs ( E, F ) on different coatings characterized via fluorescence microscopy with staining for nuclei (DAPI, Blue). The average fluorescence intensity of PLGA nanoparticles inside NIH3T3 cells ( B ), RAW264.7 cells ( D ), and ASCs ( F ) measured via ImageJ analysis of fluorescence images. Quantification was performed after 12h and 24h of incubation with PLGA nanoparticles. All data presented as mean ± SEM. (n>25 cells per group from 25 fields of view and 2 independent experiments). Two-way ANOVA with Tukey’s multiple comparison test. Symbols for statistical comparisons between indicated groups: * p ≤ 0.05; **p ≤ 0.01; *** p<0.001; or ## p<0.05 vs. all other groups. (scale bars = 40μm).

Article Snippet: Three different cell types, NIH3T3 fibroblasts (ATCC, Manassas, VA, USA, Cat. No. CRL-1658), RAW264.7 macrophages (ATCC, Manassas, VA, USA, Cat. No. TIB-71), and primary rat adipose-derived stem cells (ASCs), isolated from female 8–14 week-old Lewis rats (Charles River Laboratories, Charleston, SC, USA) were used.

Techniques: Fluorescence, Microscopy, Staining, Incubation, Comparison

Journal: ACS applied materials & interfaces

Article Title: Cellular Uptake of Nanoparticles is Regulated by Integrin-Based Adhesion to the Extracellular Matrix

doi: 10.1021/acsami.5c18030

Figure Lengend Snippet: PLGA nanoparticle uptake on NIH3T3 cells characterized via live imaging of cells for 60 min with 1 frame every 5 min via fluorescence microscope. ( A ) Average fluorescence intensity of PLGA nanoparticles inside the NIH3T3 cells measured via ImageJ analysis of fluorescence images. ( B ) Representative green PLGA fluorescence and Dil fluorescence images of NIH3T3 cells taken at the indicated time points on FN, COL, and LM coatings. All data presented as mean ± SEM (n=3–6 cells / group). Two-way ANOVA analysis with Tukey’s multiple comparison test were conducted. (scale bar = 40μm)

Article Snippet: Three different cell types, NIH3T3 fibroblasts (ATCC, Manassas, VA, USA, Cat. No. CRL-1658), RAW264.7 macrophages (ATCC, Manassas, VA, USA, Cat. No. TIB-71), and primary rat adipose-derived stem cells (ASCs), isolated from female 8–14 week-old Lewis rats (Charles River Laboratories, Charleston, SC, USA) were used.

Techniques: Imaging, Fluorescence, Microscopy, Comparison

Journal: ACS applied materials & interfaces

Article Title: Cellular Uptake of Nanoparticles is Regulated by Integrin-Based Adhesion to the Extracellular Matrix

doi: 10.1021/acsami.5c18030

Figure Lengend Snippet: Inhibition with Cytochalasin D limits focal adhesion formation and cytoskeletal arrangement of NIH3T3 cells on different coatings after 24h incubation of cells seeded with cytochalasin D. Representative fluorescence images of cytoskeletal arrangement via phalloidin staining and focal adhesion formation via paxillin staining of NIH3T3 cells on different ECM and ECM-mimetic coatings ( A ). Cells seeded on FN coatings without cytochalasin D was used as a control. PLGA nanoparticle uptake in NIH3T3 cells with or without the endocytosis pathway inhibitor chytochalasin D, characterized via fluorescence microscope (B) . Average fluorescence intensity of PLGA nanoparticles inside the NIH3T3 cells ( C ), after 12h and 24h incubation of PLGA nanoparticles with the cells seeded with/without Cytochalasin D on different coating types. All data presented as mean ± SEM (n= 2 with 15 images each sample). Two-way ANOVA analysis with Tukey’s multiple comparison test were conducted. ****p ≤ 0.0001; *** p ≤ 0.001; * p ≤ 0.05

Article Snippet: Three different cell types, NIH3T3 fibroblasts (ATCC, Manassas, VA, USA, Cat. No. CRL-1658), RAW264.7 macrophages (ATCC, Manassas, VA, USA, Cat. No. TIB-71), and primary rat adipose-derived stem cells (ASCs), isolated from female 8–14 week-old Lewis rats (Charles River Laboratories, Charleston, SC, USA) were used.

Techniques: Inhibition, Incubation, Fluorescence, Staining, Control, Microscopy, Comparison

Journal: Cell Death Discovery

Article Title: Sibiriline, a novel dual inhibitor of necroptosis and ferroptosis, prevents RIPK1 kinase activity and (phospho)lipid peroxidation as a potential therapeutic strategy

doi: 10.1038/s41420-025-02852-8

Figure Lengend Snippet: A SH-SY5Y cells were treated with either 10 µM Erastin, 5 µM RSL3, 10 µM FIN56 or 25 µM FINO 2 , and increasing concentrations of Sib up to 50 µM or until reaching maximal cellular viability or 1 µM of ferrostatin-1 (Fer-1/F1) as a control. Cell viability was determined after 24 h of treatment, using the MTS assay. The bar graph represents the mean of two replicates. B SH-SY5Y cells were co-treated for 24 h with 5 µM RSL3 and increasing concentrations of Sib or Fer-1. Cell death was estimated by the lactate dehydrogenase (LDH) release assay. Results are plotted in % of LDH release measured in cells treated with RSL3 alone (left axis, colored blue). Cell viability was evaluated by MTS reduction assay. Results obtained (colored red) were plotted as % of maximal viability with DMSO-treated cells (right axis). Data are shown as the mean +/- SEM of three replicates. C NIH3T3 cells were treated with either 1 µM RSL3, 5 ng/ml TNFα and 20 µM z-VAD.fmk (TZ) or a combination of both treatment (TZ + RSL3) and 10 µM of Sib or Nec-1f, 30 µM Nec-1s or 1 µM of Fer-1. Cell viability was evaluated by MTS reduction assay after 16 h of treatment. Data are shown as the mean ± SEM of three replicates of two independent experiments. D NIH3T3 cells were treated with 5 ng/ml TNFα, 20 µM z-VAD.fmk and 1 µM RSL3 and increasing concentrations of Sib or Nec-1f. Cell viability was determined after 16 h of treatment using an MTS assay. Data are shown as the mean ± SEM of two replicates. EC 50 values were calculated using graphpad prism software. Statistical analysis was performed using two-way ANOVA and Tukey’s multiple comparisons test using graphpad prism software. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 vs controls.

Article Snippet: Human neuroblastoma SH-SY5Y cell line, human fibrosarcoma HT1080 cell line, mouse fibroblast NIH3T3 cell line and mouse hippocampal HT-22 cell line were originally obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Control, MTS Assay, Lactate Dehydrogenase Assay, Software