Journal: Cancers

Article Title: A Double-Negative Feedback Interaction between miR-21 and PPAR-α in Clear Renal Cell Carcinoma

doi: 10.3390/cancers14030795

Figure Lengend Snippet: miR-21 and PPAR-α expressions in HK-2 and renal cancer cell lines before confluence (BC) and two days after confluence (C + 2). ( A ) Using RT-qPCR, miR-21 expression was determined in the 786-O, ACHN, RCC10, and RCC4 renal cancer cells as compared to normal HK-2 renal epithelial cells. RNU48 was used as an internal control. The values are means ± SEM and represent at least three separate experiments (* p < 0.05, ** p < 0.01, and *** p < 0.001). ( B ) Western blotting was performed on whole cell extracts. Antibodies against PPAR-α and β-actin were used.

Article Snippet: The HK-2 human proximal tubule epithelial cell line, ACHN, and 786-O human renal cancer cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Control, Western Blot

Journal: Methods in cell biology

Article Title: Analysis of primary cilia in renal tissue and cells

doi: 10.1016/bs.mcb.2019.04.008

Figure Lengend Snippet: Immortalized and primary renal epithelial cells.

Article Snippet: The balance between cilia assembly and disassembly regulates cilia length ( Mirvis, Stearns, & James Nelson, 2018 ; Spalluto, Wilson, & Hearn, 2013 ). table ft1 table-wrap mode="anchored" t5 caption a7 Cell line Description Source References M-1 Murine cortical collecting duct epithelial cells ATCC Stoos et al. (1991) IMCD-3 Murine inner medullary collecting duct epithelial cells ATCC Rauchman et al. (1993) LLC-PK1 Porcine proximal tubule epithelial cells ATCC Nielsen et al. (1998) MDCK Madin-Darby canine kidney epithelial cells ATCC Gaush et al. (1966) NHK/ADPKD Primary cortical epithelial cells from normal human kidney (NHK) or Autosomal Dominant PKD (ADPKD) KUMC Graham et al. (1977) and Reif et al. (2011) Open in a separate window Immortalized and primary renal epithelial cells. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Company Cilia structure References Acetylated-α-tubulin Sigma-Aldrich Axoneme Ishikawa et al. (2012) , Seixas et al. (2015) , Silva et al. (2018) , and Yu, Sharma, Skowronek, and Erdmann (2016) γ-tubulin Sigma-Aldrich Centrioles Breslow et al. (2013) Pericentrin Covance Centrioles Ishikawa et al. (2012) ARL13B Proteintech Ciliary membrane Seixas et al. (2015) INPP5E Proteintech Ciliary membrane Plotnikova et al. (2015) IFT52 Proteintech Axoneme Silva et al. (2018) IFT81 Proteintech Axoneme Silva et al. (2018) IFT88 Proteintech Axoneme Silva et al. (2018) IFT140 Proteintech Axoneme Silva et al. (2018) BBS2 Proteintech Axoneme Silva et al. (2018) BBS5 Proteintech Axoneme Silva et al. (2018) Open in a separate window Markers of primary cilia.

Techniques:

Journal: bioRxiv

Article Title: Development of high-affinity, single-domain protein binders for neutralizing household allergens

doi: 10.1101/2025.08.03.668213

Figure Lengend Snippet: (a) Thermostability profile of AVA-1-1-C3 as a representative protein, as determined by differential scanning fluorimetry. (b-g) Cytotoxicity of primary human cells following 48-hour exposure to AVA-1-1-C3 or an equivalent concentration of bovine serum albumin (BSA), evaluated using the CellTiter-Glo® (CTG) luminescent cell viability assay. Cell types included human epithelial keratinocytes (HEK, b), human dermal fibroblasts (HDF, c), human follicle dermal papilla cells (HFDPC, d), human renal cortical epithelial cells (HRCEPC, e), human skeletal muscle cells (HSKMC, f), and human dermal microvascular endothelial cells (HDMEC, g). No significant differences in cell viability were observed. Data in (b-g) represents mean ± s.d. and are representative of two experimental replicates.

Article Snippet: For in vitro toxicity studies, primary cells including human dermal fibroblasts (HDF, Cell Applications, Cat# 106K-05a, lot 1632), human epidermal keratinocytes (HEK, Cell Applications, Cat# 102-05a, lot 2146), human dermal microvascular endothelial cells (HDMEC, PromoCell, Cat# C-12210, lot 483Z001.3), human skeletal muscle cells (HSKMC, Cell Applications, Cat# 150K-05a, lot 3507), and human renal cortical epithelial cells (HRCEpC, Promocell, Cat# C-12660, lot 501Z019.23).

Techniques: Concentration Assay, Cell Viability Assay

Journal: Experimental and Therapeutic Medicine

Article Title: High lncRNA MEG3 expression is associated with high mortality rates in patients with sepsis and increased lipopolysaccharide-induced renal epithelial cell and cardiomyocyte apoptosis

doi: 10.3892/etm.2019.8049

Figure Lengend Snippet: LncRNA MEG3 regulates renal epithelial cell and cardiomyocyte apoptosis following LPS treatment. LncRNA MEG3 overexpression and knockdown were confirmed. Following LPS treatment, LncRNA MEG3 overexpression resulted in significantly increased apoptosis, whereas lncRNA MEG3 knockdown by siRNA significantly inhibited apoptosis in (A) renal epithelial cells and (B) AC12 cardiomyocytes. *P<0.05. LncRNA, long non-coding RNA; MEG3, maternally expressed 3; LPS, lipopolysaccharide.

Article Snippet: Human primary renal mixed epithelial cells (ATCC ® PCS-400-012TM) were purchased from American Type Culture Collection.

Techniques: Over Expression, Knockdown

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Negative regulatory responses to metabolically triggered inflammation impair renal epithelial immunity in diabetes mellitus

doi: 10.1007/s00109-012-0969-x

Figure Lengend Snippet: Renal tubular epithelial cells, but not mesangial cells, modulate overt inflammation via negative feedback mechanisms that impair immunity. a Human renal primary tubular epithelial cells ( RPTEC ) and mesangial cells ( NHMC ; Lonza) were cultured with diabetes-relevant concentrations of cytokines (TNF-α, 50 pg/ml; IL-6, 20 pg/ml; and IL-1β, 20 pg/ml), high glucose ( HG ; 20 mM) or methylglyoxal ( MG ; 10 μM) for 3 h or 5 days (120 h). Protein lysates from each condition ( n = 4) were prepared for western blot analysis using the indicated antibodies ( n ≥ 3 for each antibody). b RPTEC were first exposed to a combination of cytokines, high glucose, and methylglyoxal at concentrations stated in ( a ) for 5 days. The cells were further cultured in fresh medium in the presence of either diphenyleneiodonium chloride ( DPI ; 10 μM) or combined with Gö6983 (10 nM) for 1 day before challenge with lipopolysaccharide ( LPS ; 1 μg/ml) for 2 h. Protein lysates ( n = 4 for each condition) were analyzed by western blotting (≥3 immunoblots for each antibody). c RPTEC cultured in the combined presence of cytokines, HG and MG at the aforementioned concentrations for 5 days were challenged with a higher dose of LPS (10 μg/ml) for the indicated time points ( n = 4 for each time point). Minus 1 h denotes cells that were not exposed to LPS but were lysed for protein isolation one hour before similarly cultured cells were challenged with LPS. Protein lysates from each condition were prepared for western blotting as described (Figs. and legends). d RPTEC and NHMC were treated as follows before harvesting for protein and total RNA isolations ( n = 4 for each condition). I Untreated control cells; II incubated with cytokines (TNF-α, 50 pg/ml; IL-6, 20 pg/ml; and IL-1β, 20 pg/ml), HG (20 mM), and MG (10 μM) for 5 days; III incubated with MG (10 μM) only for 5 days; IV challenged with LPS (10 μg/ml), lipoteichoic acid ( LTA ; 10 μg/ml), and poly(I:C) (10 μg/ml) for 2 h; V treated as in ( II ) before returning to normal culture medium for 1 day and subsequently challenged with microbial stimulants as in ( IV ); VI cultured as in ( II ) before treatment with DPI (10 μM) and Gö6983 (10 nM) for 1 day in fresh culture medium; VII cultured as in ( VI ) before challenge as in ( IV ). Protein lysates were assayed using quantitative protein arrays. e Results of qPCR array plotted as fold change (RPTEC of each condition cf. untreated RPTEC). Genes with similar functions or involvement in the same biological process were grouped. A Diabetes related, B inflammatory pathways, C antioxidant system, D negative immune regulators, E interleukin and cytokine receptors, F TLR signaling, and G antimicrobial molecules and adaptive immunity. Roman numerals ( I–V ) indicate the type of treatment. I RPTEC incubated with cytokines (TNF-α, 50 pg/ml; IL-6, 20 pg/ml; and IL-1β, 20 pg/ml), HG (20 mM), and MG (10 μM) for 5 days; II RPTEC challenged with LPS (10 μg/ml), LTA (10 μg/ml) and poly I/C (10 μg/ml) for 2 h; III RPTEC treated as in ( I ), returned to normal culture medium for 1 day and then challenged with microbial analogs as in ( II ); IV RPTEC cultured as in ( I ) before combined treatment with DPI (10 μM) and Gö6983 (10 nM) for 1 day in fresh culture medium; V RPTEC cultured as in ( IV ) before being challenged as in ( II ). The qPCR array experiment for each of the conditions was performed twice

Article Snippet: Clonetics® human primary renal tubular epithelial cells (RPTEC) and mesangial cells (NHMC), purchased from Lonza BioSciences, were cultured in REGMTM Renal Epithelial Cell Growth Medium and NHMC in MsGMTM Mesangial Cell Growth Medium, respectively.

Techniques: Cell Culture, Western Blot, Isolation, Control, Incubation