Journal: Scientific Reports

Article Title: 5T4-specific chimeric antigen receptor modification promotes the immune efficacy of cytokine-induced killer cells against nasopharyngeal carcinoma stem cell-like cells

doi: 10.1038/s41598-017-04756-9

Figure Lengend Snippet: 5T4-28Z CAR gene modification promotes the anti-NPC activity of CIK cells in a 5T4-dependent manner. Target cells consisted of 5T4 low-expressing S18 and S26 cells and their spheroid cells with an intermediate or high level of 5T4 expression. Effector cells included 5T4-28Z-CIK, 5T4-28Z-CIK (NKG2D-), C-28Z-CIK and NT-CIK cells. ( a–d ) Briefly, 1 × 10 4 target cells per well were co-cultured with different effector cells at E/T ratios of 10/1, 5/1 and 2.5/1 for 4 h. The cytotoxicity of effector cells was examined with LDH release assays. Values in the line graphs represent the mean ± SD of three parallel wells. ( e ) Then, 2 × 10 4 target cells per well were co-incubated with 1 × 10 5 effector cells per well for 24 h. Effector cells cultured alone in medium served as the negative control. IFN-γ production of the effector cells was assessed using ELISAs. Values in the histogram represent the mean ± SD of three parallel wells. ( f ) In addition, 2 × 10 5 target cells per well were co-cultured with 1 × 10 6 effector cells per well for 5 h in the presence of 1 × Protein Transport Inhibitor Cocktail and anti-human CD107α-APC antibody or IgG isotype control. Effector cells cultured alone served as the negative control. Degranulation of effector cells was evaluated using FACS. Values presented in the histogram represent the mean ± SD of triplicate samples. ( g ) S26 cells or S26 spheroids labelled with CM-Dil were co-cultured with 5T4-28Z-CIK cells and monitored using an inverted fluorescence microscope with climate control. Images captured intermittently are displayed. The results are representative of at least three independent experiments. * indicates p < 0.05 (one-way ANOVA).

Article Snippet: Next, 50 μl of supernatant per well was collected to measure LDH release using a cytotoxicity LDH Assay Kit-WST ® (Dojindo) according to the manufacturer’s specifications.

Techniques: Modification, Activity Assay, Expressing, Cell Culture, Incubation, Negative Control, Control, Fluorescence, Microscopy

Journal: The Korean Journal of Internal Medicine

Article Title: Angiotensin III increases monocyte chemoattractant protein-1 expression in cultured human proximal tubular epithelial cells

doi: 10.3904/kjim.2016.31.1.116

Figure Lengend Snippet: Angiotensin III (Ang III)-stimulated monocyte chemoattractant protein-1 (MCP-1) production in HK-2 cells. Cells were incubated for the indicated times in the presence or absence of Ang III. MCP-1 protein in culture medium was quantified by enzyme-linked immunosorbent assay. Results are shown as mean ± SEM from six independent experiments. Lactate dehydrogenase (LDH) release from Ang II-, Ang III-, or losartan-treated cells. LDH release is expressed as percentage of maximal LDH release induced by 1% Triton X-100 for 48 hours. a p < 0.05 vs. 8 hours MCP-1 level, b p < 0.05 vs, control cells.

Article Snippet: HK-2 cell viability was assessed by measuring lactate dehydrogenase (LDH) release into the culture medium using a LDH cytotoxicity detection kit according to the manufacturer’s protocol (Takara Biomedical, Kyoto, Japan).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Korean Journal of Internal Medicine

Article Title: Angiotensin III increases monocyte chemoattractant protein-1 expression in cultured human proximal tubular epithelial cells

doi: 10.3904/kjim.2016.31.1.116

Figure Lengend Snippet: Lactate dehydrogenase (LDH) release from angiotensin (Ang) II-, Ang III-, or losartan-treated cells. LDH release is expressed as percentage of maximal LDH release induced by 1% Triton X-100 for 48 hours.

Article Snippet: HK-2 cell viability was assessed by measuring lactate dehydrogenase (LDH) release into the culture medium using a LDH cytotoxicity detection kit according to the manufacturer’s protocol (Takara Biomedical, Kyoto, Japan).

Techniques:

Journal: BMC oral health

Article Title: Keratinocyte necroptosis promotes the progression of radiation-induced oral mucositis.

doi: 10.1186/s12903-025-06278-7

Figure Lengend Snippet: Fig. 4 Pretreatment with RIPK3 inhibitor or MLKL inhibitor reduces radiation-induced cell death and inflammation. (A) Lactate dehydrogenase (LDH) release was measured in HaCaT cells cultured for 2.5 days post-irradiation with a dose of 12 Gy, in the presence or absence of the RIPK3 inhibitor GSK’872 or the MLKL inhibitor GW806742X. (B) Flow cytometry analysis of HaCaT cells stained with propidium iodide (PI) and annexin V, and (C) quantification of double-positive cells (PI and annexin V staining). (D) Representative fluorescent images of Hoechst 33342 (blue) and PI (red) double staining. Scale bar: 100 μm. (E, F) The mRNA expression levels of inflammatory cytokines by qRT-PCR, including IL-1β, IL-6, and TNF-α. Results are shown as ± SD means of three replicates from three independent experiments. Statistical significance was determined using one-way ANOVA and LSD post hoc tests. *P < 0.05, **P < 0.01 and ***P < 0.001

Article Snippet: Cytotoxicity was evaluated using the Lactate Dehydrogenase (LDH) Assay Kit (Beyotime).

Techniques: Cell Culture, Irradiation, Flow Cytometry, Staining, Double Staining, Expressing, Quantitative RT-PCR