Journal: ACS applied materials & interfaces

Article Title: Enhanced Cell Proliferation and Maturation Using Carboxylated Bacterial Nanocellulose Scaffolds for 3D Cell Culture.

doi: 10.1021/acsami.4c22475

Figure Lengend Snippet: Figure 5. Biocompatibility of BNC and BNC−COOH hydrogels. (A) Cell viability of 3T3-L1 fibroblasts in BNC and BNC−COOH hydrogels over 7, 14, and 21 days. (B) Confocal microscopy micrographs of the 3T3-L1 cells cultured in BNC and BNC−COOH hydrogels at 21 days (scale bars represent 40 μm). Cells were stained with resorufin (pink color), while bacterial nanofibers were stained with calcofluor (blue color). (C) Mean fluorescence intensity of the DCFDA (shown in green in D) for quantifying ROS production in cells grown in hydrogels. (D) ROS production in 3T3-L1 cells at 21 days. Cells are green in color due to the fluorescence of DCFDA, and the bacterial nanofibers are blue due to the staining with calcofluor (scale bars represent 20 μm). The bars represent the mean ± standard deviation. Statistical significance is as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The fibroblast cell line (NIH-3T3-L1) was obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, USA).

Techniques: Confocal Microscopy, Cell Culture, Staining, Fluorescence, Standard Deviation

Journal: Polymers

Article Title: A Novel Approach for the Manufacturing of Gelatin-Methacryloyl

doi: 10.3390/polym14245424

Figure Lengend Snippet: Cell viability of NIH 3T3 fibroblasts on gelatin methacryloyl (GelMA) coated well plates. GelMA produced similar to the method of Shirahama at 50 ∘ C (50C100MA) was used as a control in comparison with GelMA produced by the novel approach presented in this study, i.e., at room temperature (RT100MA). The viability of cells seeded onto both hydrogels samples did not differ significantly after one and three days.

Article Snippet: NIH 3T3 mouse fibroblasts (CLS Cell Lines Service, Eppelheim, Germany) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, high glucose, GlutaMAXTM) supplemented with 10% ( v/v ) FBS (fetal bovine serum), 50 U mL −1 penicillin, and 50 μg mL −1 streptomycin.

Techniques: Produced, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Absolute Protein Amounts and Relative Abundance of Volume-regulated Anion Channel (VRAC) LRRC8 Subunits in Cells and Tissues Revealed by Quantitative Immunoblotting

doi: 10.3390/ijms20235879

Figure Lengend Snippet: Quantification of LRRC8 protein amounts in murine cell lines. ( A , B ) Two replicates of whole-cell protein preparations from wild-type C2C12 (A) and 3T3 (B) cells (WT-1 and WT-2) and from a LRRC8A-deficient C2C12 and 3T3 line (KO), with 60 µg/lane, were separated by SDS-PAGE. Each blot was loaded with a dilution of recombinant GST fusion protein to calibrate for the respective antibody signal. The size of the LRRC8 proteins, as judged from the LRRC8A KO control or from comparison to data from human cells lacking all five LRRC8 proteins ( , ), is indicated. The blots are representative for three independent experiments. ( C , D ) Quantification of LRRC8A-E in C2C12 (C) and 3T3 ((D) cells from three independent blots with two lysates each. Data represent the mean from six lysates ± SD. *** p < 0.001, n.s. = not significant, compared with LRRC8A using one-way analysis of variance (ANOVA) with Bonferroni’s post hoc test.

Article Snippet: 3T3 mouse fibroblast cell line (American Type Culture Collection, Manassas, VA, USA; ATCC CL-173) and C2C12 mouse skeletal muscle myoblasts (American Type Culture Collection, Manassas, VA, USA) were kindly provided by P. Knaus, Freie Universität Berlin, Germany.

Techniques: SDS Page, Recombinant, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Absolute Protein Amounts and Relative Abundance of Volume-regulated Anion Channel (VRAC) LRRC8 Subunits in Cells and Tissues Revealed by Quantitative Immunoblotting

doi: 10.3390/ijms20235879

Figure Lengend Snippet: Quantification of LRRC8 protein amounts in co-immunoprecipitation with LRRC8A. ( A , B ) LRRC8A co-precipitated LRRC8B-E in immunoprecipitations with an LRRC8A antibody from C2C12 (A) and LRRC8B-D from 3T3 cell lysates (B), but not from the respective LRRC8A-deficient cells. The Na,K-ATPase, tested as negative control, was not co-precipitated. Lysate equivalent to 25% of input was loaded as reference (input). Each blot for LRRC8A-E was loaded with a dilution of recombinant GST fusion protein to calibrate for the respective antibody signal. ( C , D ) Quantification of precipitated LRRC8A-E in C2C12 (C) and 3T3 (D) cells, per µg of total protein subjected to the immunoprecipitation. Data represent mean ± SD from three independent experiments. * p < 0.05, *** p < 0.001, n.s. = not significant, compared with LRRC8A using one-way ANOVA with Bonferroni’s post hoc test.

Article Snippet: 3T3 mouse fibroblast cell line (American Type Culture Collection, Manassas, VA, USA; ATCC CL-173) and C2C12 mouse skeletal muscle myoblasts (American Type Culture Collection, Manassas, VA, USA) were kindly provided by P. Knaus, Freie Universität Berlin, Germany.

Techniques: Immunoprecipitation, Negative Control, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: Absolute Protein Amounts and Relative Abundance of Volume-regulated Anion Channel (VRAC) LRRC8 Subunits in Cells and Tissues Revealed by Quantitative Immunoblotting

doi: 10.3390/ijms20235879

Figure Lengend Snippet: Immunoblot against the five LRRC8 proteins in lysates of different cell lines (and LRRC8A-KO of C2C12 and 3T3) and organs. ( A ) Equal amounts (60 µg protein/lane) of cell and tissue lysates were separated by SDS-PAGE, β-actin (not expressed in heart ) served as a loading control. The size of the LRRC8 proteins, as judged from the LRRC8A KO control or from comparison to data from human cells lacking all five LRRC8 proteins ( , ), is indicated. ( B ) Equal loading of wells was additionally verified by Ponceau staining after protein transfer and by probing for GM130 and GAPDH as loading controls for each immunoblot (examples shown).

Article Snippet: 3T3 mouse fibroblast cell line (American Type Culture Collection, Manassas, VA, USA; ATCC CL-173) and C2C12 mouse skeletal muscle myoblasts (American Type Culture Collection, Manassas, VA, USA) were kindly provided by P. Knaus, Freie Universität Berlin, Germany.

Techniques: Western Blot, SDS Page, Control, Comparison, Staining