Journal:

Article Title: Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract

doi: 10.1128/IAI.72.6.3171-3178.2004

Figure Lengend Snippet: Bladder cells take up AOAH precursor. (A) Uptake of pro-AOAH by T24 cells after incubation with pro-AOAH-containing medium for 5 h in the presence or absence of 10 mM M6P, G6P, or mannose. Washed cells were lysed and AOAH protein was assayed by ELISA. Each bar shows the mean and standard error of data from four independent experiments. (B) AOAH specific activity (activity/nanograms of protein) in T24 cell lysates. Compared to the pro-AOAH added in the medium (AOAH Source), cell-associated AOAH had a much greater specific activity, reflecting its activation by the T24 cells. M6P inhibited AOAH binding (A) but did not prevent the activation of the cell-associated AOAH (B).

Article Snippet: LLC-PK1 porcine proximal tubule cells (American Type Culture Collection [ATCC], CL-101), T24 human bladder cells (ATCC, HTB-4), and AOAH-transfected and untransfected BHK570 cells ( 31 ) were cultured in low-glucose Dulbecco modified Eagle medium (Invitrogen, Carlsbad, Calif.), Vitacell McCoy's 5a medium (ATCC), or DMEM with glutamine and 4.5 g of glucose (Fisher Scientific)/liter, respectively.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Activity Assay, Activation Assay, Binding Assay

Journal:

Article Title: Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract

doi: 10.1128/IAI.72.6.3171-3178.2004

Figure Lengend Snippet: AOAH confers LPS-deacylating activity to bladder cells. T24 cells were allowed to take up AOAH for 5 h, washed, and then incubated with [3H]LPS (125 ng/ml) for the times indicated. Control cells were incubated with medium that did not contain AOAH. Whereas control and AOAH-containing cells took up similar amounts of [3H]LPS (A), only the AOAH-containing cells removed 3H-labeled fatty acids from the LPS backbone (B). The data represent combined results of three separate experiments; the error bars represent one standard error of the mean. Solid squares and bars, T24 cells with AOAH; open circles and bars, control T24 cells.

Article Snippet: LLC-PK1 porcine proximal tubule cells (American Type Culture Collection [ATCC], CL-101), T24 human bladder cells (ATCC, HTB-4), and AOAH-transfected and untransfected BHK570 cells ( 31 ) were cultured in low-glucose Dulbecco modified Eagle medium (Invitrogen, Carlsbad, Calif.), Vitacell McCoy's 5a medium (ATCC), or DMEM with glutamine and 4.5 g of glucose (Fisher Scientific)/liter, respectively.

Techniques: Activity Assay, Incubation, Control, Labeling

Journal: BMC Infectious Diseases

Article Title: Use of ceragenins as a potential treatment for urinary tract infections

doi: 10.1186/s12879-019-3994-3

Figure Lengend Snippet: Cell viability of T24 bladder cancer cell line after treatment with LL-37 (filled circles), doxycyclinum (filled squares), CSA-13 (filled diamonds), CSA-131 (filled triangles), LL-37 with doxycyclinum (empty circles), LL-37 with CSA-13 (empty squares) and LL-37 with CSA-131 (empty triangles)

Article Snippet: Human bladder epithelial cell line T24 (ATCC® HTB 4TM), purchased from American Type Culture Collection (ATCC), was cultured at 37 °C with 5% CO 2 in McCoy’s 5A medium (ATCC, catalog no. 30–2007) supplemented with 10% fetal bovine serum (FBS, ATCC, catalog no. 30–2020).

Techniques:

Journal: BMC Infectious Diseases

Article Title: Use of ceragenins as a potential treatment for urinary tract infections

doi: 10.1186/s12879-019-3994-3

Figure Lengend Snippet: Intracellular killing efficacies of LL-37 (filled circles), doxycyclinum (filled squares), CSA-13 (filled diamonds), CSA-131 (filled triangles) and combination of LL-37 with doxycyclinum (empty circles), LL-37 with CSA-13 (empty squares) and LL-37 with CSA-131 (empty triangles) against four clinical strains of E. coli* within T24 cells (Panels a - d ). The concentrations of used antibacterial agents were 5 and 10 μM; incubation time was 2 h

Article Snippet: Human bladder epithelial cell line T24 (ATCC® HTB 4TM), purchased from American Type Culture Collection (ATCC), was cultured at 37 °C with 5% CO 2 in McCoy’s 5A medium (ATCC, catalog no. 30–2007) supplemented with 10% fetal bovine serum (FBS, ATCC, catalog no. 30–2020).

Techniques: Incubation

Journal: Nature Communications

Article Title: Chromatin-associated degradation is defined by UBXN-3/FAF1 to safeguard DNA replication fork progression

doi: 10.1038/ncomms10612

Figure Lengend Snippet: ( a ) Western blot analysis of replication checkpoint activation by phosho-specific antibodies in HEK293T cells transfected with indicated siRNAs. ( b ) Schematic illustration of DNA replication fibre analysis—(a) Indicates ongoing replication (red and green arrows), (b) stalled replication forks (red arrows only), (c) newly fired origins (green arrows only) and (d) terminated/converged forks. Sites of DNA replication initiation (I) or termination (T) are labelled, respectively. ( c ) Western blot analysis of T24 cell lysates transfected with indicated siRNAs. Efficiency of siRNA-mediated depletion is shown for FAF1 protein levels. ( d , e ) Representative images of microscopic analysis of DNA replication fibres by molecular combing in T24 cells after indicated siRNA transfection. CldU (first pulse) incorporation is shown in red, incorporated IdU (second pulse) in green. Quantification of replicated DNA tract length after indicated siRNA transfection in T24 cells. Tract length was determined for 100 individual forks per condition and experiment. The experiment was repeated twice. Whisker box plots show mean values and data within the 10–90 percentile. ( f – i ) Representative images of microscopic analysis of DNA replication fibres by molecular combing in U2OS cells after indicated siRNA transfection. Quantification of replicated DNA tract length, stalled replication forks and newly fired dormant origins after indicated siRNA transfection in U2OS cells. Tract length was determined for 100 forks per condition and experiment. Fork stalling and origin firing was quantified for 400 forks per condition and experiment. The experiment was repeated in three replicates. ( j ) Western blot analysis of control HA and HA-FAF1 co-IPs. Nuclear lysates were prepared from HEK cells expressing the p97-E587Q variant. ( k ) HEK cells were depleted for control or FAF1 by siRNA. CDT-1 and PCNA levels were monitored at indicated time points after CHX treatment by western blot analysis. Scale bar, 5 μm. Data show mean values. Error bars represent s.d.. The single asterisk indicates a P value of ≤0.05, the double asterisk ≤0.001 and the triple asterisk indicate P values of ≤0.0001.

Article Snippet: Human osteosarcoma (U2OS) cells, human embryonic kidney 293 (HEK293) cells and human bladder cancer (T24) cells (cell lines were purchased at ATCC, UK (CRL-1573, HTB-96 and HTB-4]) were cultured in six-well plates, 60- or 100-mm dishes in DMEM supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin in a 5% CO 2 atmosphere at 37 °C.

Techniques: Western Blot, Activation Assay, Transfection, Whisker Assay, Control, Expressing, Variant Assay