bj 5ta immortalized fibroblast cell lines  (ATCC)

Journal: bioRxiv

Article Title: Eukaryotic Initiation Factor 5B (eIF5B)-Driven Translational Control Impacts Oral Squamous Cell Carcinoma Pathophysiology

doi: 10.64898/2026.01.30.702967

Figure Lengend Snippet: BJ-5ta cells were transfected with either a non-specific control siRNA (siC) or an eIF5B-specific siRNA pool (si-eIF5B). (A) Following 24 h of transfection, cells were treated with either vehicle control or TRAIL (100 ng/mL). Cell viability was assessed after an additional 72 h using alamarBlue TM assays. (B) Annexin V and 7-AAD staining and flow cytometric analysis of transfected cells. The top two panels, siC and siC + TRAIL, the bottom two panels, si5B and si5B + TRAIL, were harvested at 96 h post-transfection and treated with Annexin V + 7AAD. Contour plots are divided into quadrants Live (lower left; cells staining negative for both 7-AAD and annexin V), Q1 (upper left; cells staining positive for 7-AAD but negative for annexin V), Early apoptosis (lower right; staining annexin V positive and 7-AAD negative), and Late apoptosis (upper right; staining positive for both 7-AAD and annexin V). (C) Representative immunoblots probing for eIF5B, XIAP, Bcl-xL, cIAP1, cFLIPs, DR 4, DR 5, and β-actin (internal control) are shown. ( D) Control and eIF5B-depleted BJ-5ta cells were treated with the 4EGI-1 inhibitor to inhibit cap-dependent translation. Levels of XIAP were monitored under control and the 4EGI-1 treatment conditions. (E) eIF5B-bound RNA immunoprecipitations (RIPs) followed by qRT-PCR measurements of input-normalized tRNAiMet levels from BJ-5ta vs UMSCC-29. Data are presented as mean ± SEM from three independent biological replicates, with statistical significance indicated as follows: *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: CAL-33 and BJ-5ta (immortalized fibroblast) cell lines were obtained from American Type Culture Collection (ATCC; Virginia, USA), and UMSCC-1 and UMSCC-29 were obtained from the University of Michigan (Michigan, USA).

Techniques: Transfection, Control, Staining, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Eukaryotic Initiation Factor 5B (eIF5B)-Driven Translational Control Impacts Oral Squamous Cell Carcinoma Pathophysiology

doi: 10.64898/2026.01.30.702967

Figure Lengend Snippet: BrdU incorporation assay was used to measure cell proliferation. Quantification shows the percentage of BrdU-labeled cells in control (siC) and si5B-treated groups. Data are presented as mean ± SEM; ***p < 0.001. (A) UMSCC-29 (B) BJ-5ta (C) Collagen-based invasion assay was used to evaluate cell invasiveness in control (siC) and si5B-treated groups. Cells were seeded onto collagen matrices and allowed to invade for a defined period (48-72 hours). The number of invading cells was quantified and expressed as the mean ± SEM. Microscopy images of invading cells (C; left panel) , quantification (C; right panel) . Phase contrast microscopy images showing wound area in siC/si5B-treated OSCC cells at the time of scratch introduction (T = 0 h) & at T = 30 h wound closure (c; top panel) . Quantification of the percentage of wound area covered after 30 hours, normalized to wound area at the time of scratch introduction (c; lower panel) . (E) Representative images of the endothelial HUVEC cells tube formation assay treated with conditioned media from siC- or si5B-transfected OSCC. ImageJ angiogenesis macro software quantified Junctions, segments, and nodes. OSCC cells were transfected with either control siRNA (siC) or eIF5B-specific siRNA (si5B) and incubated for 96 hours. (F) The conditioned media for the control and eIF5B-depleted OSCC cells were analyzed for angiogenic biomarkers (lower panel) using EVE Technologies. (G) Representative images of immunoblots quantified probing for eIF5B, p-EGFR, EGFR, p-ERK, ERK, p-NF-kB, NF-kB, HIF1α, VEGFA, and β-actin (internal control). Data are presented as mean ± SEM from three independent biological replicates, with statistical significance indicated as follows: *, p < 0.05; **, p < 0.01; ***, p < 0.001, and ****, p< 0.0001.

Article Snippet: CAL-33 and BJ-5ta (immortalized fibroblast) cell lines were obtained from American Type Culture Collection (ATCC; Virginia, USA), and UMSCC-1 and UMSCC-29 were obtained from the University of Michigan (Michigan, USA).

Techniques: BrdU Incorporation Assay, Labeling, Control, Invasion Assay, Microscopy, Tube Formation Assay, Transfection, Software, Incubation, Western Blot

Journal: bioRxiv

Article Title: Eukaryotic Initiation Factor 5B (eIF5B)-Driven Translational Control Impacts Oral Squamous Cell Carcinoma Pathophysiology

doi: 10.64898/2026.01.30.702967

Figure Lengend Snippet: BJ-5ta cells were transfected with either control siRNA (siC) or eIF5B-specific siRNA (si5B) and incubated for 96 hours. TRAIL (100 ng/mL) treatment was performed for 4 hours, then the media was collected. The conditioned media for the control and eIF5B-depleted BT5Ta cells were analyzed for angiogenic biomarkers using EVE Technologies. The level of distinct angiogenic biomarkers was decreased in the spent media of eIF5B-depleted BJ-5ta cells. Data are expressed as mean ± SEM for three biological replicates. Statistical significance is indicated as *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: CAL-33 and BJ-5ta (immortalized fibroblast) cell lines were obtained from American Type Culture Collection (ATCC; Virginia, USA), and UMSCC-1 and UMSCC-29 were obtained from the University of Michigan (Michigan, USA).

Techniques: Transfection, Control, Incubation

Journal: Pathogens

Article Title: A Major Facilitator Superfamily Transporter Is Critical for the Metabolism and Biogenesis of the Apicoplast

doi: 10.3390/pathogens14080763

Figure Lengend Snippet: TgApMFS1 is required for the survival of parasites. ( a ) Plaque assays were performed by infecting BJ-5ta cells with TgApMFS1 loxp parasites for 8 days in the presence or absence of rapamycin. WT, wild type; ( b ) Replication of TgApMFS1 loxp parasites. The parasites were treated with or without rapamycin for 72 h, then inoculated into BJ-5ta cells in 12-well plates and allowed to grow with or without rapamycin for 24 h. The average number of parasites per vacuole was determined. The data were analyzed by a two-way ANOVA; **** p < 0.0001.

Article Snippet: The parasites were cultured on the hTERT-immortalized foreskin fibroblast cell line BJ-5ta (ATCC ® CRL-4001) in Dulbecco’s Modified Eagle Medium (DMEM) + 10% Fetal Bovine Serum (FBS).

Techniques:

Journal: Pathogens

Article Title: A Major Facilitator Superfamily Transporter Is Critical for the Metabolism and Biogenesis of the Apicoplast

doi: 10.3390/pathogens14080763

Figure Lengend Snippet: Depletion of TgApMFS1 disturbs fatty acid synthesis in the parasites. ( a ) GC-MS analysis of fatty acids in parasites lacking TgApMFS1. BJ-5ta cells were cultured with 13C-glucose for 6 h, inoculated with parasites, and then cultured with rapamycin for an additional 48 h. The egressed parasites were collected, purified using a 5 µm filter, and analyzed by GC-MS. The wild type parasites cultured with rapamycin were used as control in this study; **** p < 0.0001, * p = 0.0136, * p = 0.035; ( b ) Mass isotopologue distributions (MID) of fatty acid (FA) (C14:0) from 13 C-glucose in the parasites lacking TgApMFS1. The x -axis indicates the number of 13 C atoms in each Fatty Acid Methyl Ester (FAME), where ‘m0′ indicates the monoisotopic mass containing no 13 C atoms, while ‘mX’ represents that mass with ‘X’ 13C atoms incorporated. **** p < 0.0001, **** p < 0.0001, * p = 0.0143, **** p < 0.0001, **** p < 0.0001, **** p < 0.0001, **** p < 0.0001, **** p < 0.0001, **** p < 0.0001, **** p < 0.0001, * p = 0.013.

Article Snippet: The parasites were cultured on the hTERT-immortalized foreskin fibroblast cell line BJ-5ta (ATCC ® CRL-4001) in Dulbecco’s Modified Eagle Medium (DMEM) + 10% Fetal Bovine Serum (FBS).

Techniques: Gas Chromatography-Mass Spectrometry, Cell Culture, Purification, Control