Journal: Cell Proliferation

Article Title: Haematopoietic lineage‐committed bone marrow cells, but not cloned cultured mesenchymal stem cells, contribute to regeneration of renal tubular epithelium after HgCl 2 ‐induced acute tubular injury

doi: 10.1111/j.1365-2184.2008.00545.x

Figure Lengend Snippet: Examples of engraftment of haematopoietic lineage marrow cells (HLMCs) and mesenchymal stem cells at 3 days after HgCl2. (a) GFP‐positive control shows proximal tubular cells (PHA‐L, blue colour) co‐stained for GFP (brown colour). (b) GFP‐negative control (wild type) showing no detection of GFP. (c) BMT mouse treated with HgCl2 demonstrating GFP‐positive proximal tubular epithelial cells. Dashed black arrows indicate PHA‐L stained donor HLMC‐derived tubular cells. (d) Male positive control; Y chromosomes were detected by indirect FISH, black arrows point to Y chromosomes (brown dot) in proximal tubular cells histochemically stained with PHA‐L (red colour). (e) Female control showing the lack of Y chromosome detection within proximal tubular cells histochemically stained with PHA‐L. (f) Scattered Y chromosomes (brown dot) in renal interstitial area, not in proximal tubular cells. (g) High magnification of yellow box in (f); a–f, ×400; g, ×600. BMT, bone marrow transplantation; FISH, fluorescence in situ hybridization; PHA‐L, Phaseolus vulgaris leucoagglutinin.

Article Snippet: After GFP staining, tissue sections were microwaved in 10 m m tri‐sodium citrate (pH 6.0) for 10 min and biotin was blocked using a biotin blocking kit (DAKO), and then incubated for 45 min with biotinylated Phaseolus vulgaris leucoagglutinin (PHA‐L) 1/100 (B‐1115, Vector Laboratories, Burlingame, CA, USA) to detect proximal convoluted tubules, or biotinylated peanut agglutinin (PNA) 1/800 (B‐1075, Vector Laboratories) to detect distal convoluted tubules.

Techniques: Positive Control, Staining, Negative Control, Derivative Assay, Transplantation Assay, Fluorescence, In Situ Hybridization

Journal: Cell Proliferation

Article Title: Haematopoietic lineage‐committed bone marrow cells, but not cloned cultured mesenchymal stem cells, contribute to regeneration of renal tubular epithelium after HgCl 2 ‐induced acute tubular injury

doi: 10.1111/j.1365-2184.2008.00545.x

Figure Lengend Snippet: (a) Changes in the abundance of GFP‐positive cells within the PHA‐L stained cell population in control mice and mice treated with HgCl2 (n = 5 per treatment time point). *P < 0.05 versus the same group at day 0; +P < 0.05 versus group CON at the corresponding time point. The percentages of GFP‐positive cells were adjusted based on a correction factor derived from the actual cell count of 41% of PHA‐L stained cells being GFP‐positive in GFP donor mice. (b) Changes in the 3H‐LI of PHA‐L stained proximal tubular cells of: combined indigenous and donor HLMC (left panel); indigenous origin (central panel); donor HLMC origin (right panel). n = 5 per group. *P < 0.05 versus the same group at day 0; +P < 0.05 versus group CON at the corresponding time point. (c–g) Examples of chimerism and proliferation (3H‐thymidine labelling) of GFP+ HLMC‐derived PHA‐L‐stained cells at 3 days after HgCl2. (c) Black arrowheads indicate PHA‐L stained (blue colour in tubular cell apical membrane) donor GFP+ HLMC‐derived tubular cells (brown colour in cytoplasm) under bright field, and black arrows indicate 3H‐thymidine labelling of PHA‐L‐stained HLMC‐derived tubular cells, ×500. (d) The same field under dark field, white arrowheads point to silver grains (3H‐thymidine labelling) and white arrows indicate 3H‐thymidine labelling of PHA‐L‐stained HLMC‐derived tubular cells (×500). (e) Bright field and (f) dark field are the higher magnification of yellow box area in (c). (g) The images in (e) and (f) were combined to help to show silver grains over cells that are donor GFP+ HLMC‐derived PHA‐L‐stained tubular cells. GFP, green fluorescent protein; HLMC, haematopoietic lineage marrow cell; PHA‐L, Phaseolus vulgaris leucoagglutinin.

Article Snippet: After GFP staining, tissue sections were microwaved in 10 m m tri‐sodium citrate (pH 6.0) for 10 min and biotin was blocked using a biotin blocking kit (DAKO), and then incubated for 45 min with biotinylated Phaseolus vulgaris leucoagglutinin (PHA‐L) 1/100 (B‐1115, Vector Laboratories, Burlingame, CA, USA) to detect proximal convoluted tubules, or biotinylated peanut agglutinin (PNA) 1/800 (B‐1075, Vector Laboratories) to detect distal convoluted tubules.

Techniques: Staining, Derivative Assay, Cell Counting

Journal: Hypertension

Article Title: Downregulation of Renal TRPM7 and Increased Inflammation and Fibrosis in Aldosterone-Infused Mice

doi: 10.1161/hypertensionaha.107.100339

Figure Lengend Snippet: Figure 2. Immunohistochemical images demonstrate renal IL-6 content in the 4 groups. IL-6 is demonstrated as brown staining. IL-6 content was increased in the aldosterone group vs other groups. Magnesium reduced IL-6 content in aldosterone-treated mice. Images are representative of 8 to 9 mice in each group. Insert is a representative image from another section.

Article Snippet: For direct immunohistochemistry, sections were incubated with 3 % H2O2 and a Pierce solution to block endogenous peroxidase and biotin, respectively, followed by overnight incubation (humidified box, 4°C) with a biotinated anti-goat interleukin-6 (IL-6) monoclonal antibody (Santa Cruz).

Techniques: Immunohistochemical staining, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-4 induces apoptosis in A549 lung adenocarcinoma cells: evidence for the pivotal role of 15-hydroxyeicosatetraenoic acid binding to activated peroxisome proliferator-activated receptor gamma transcription factor.

doi: 10.4049/jimmunol.170.2.887

Figure Lengend Snippet: FIGURE 2. A, IL-4-induced apoptosis is mimicked by 15(S)-HETE and 15-PGJ2, but is inhibited by inhibitors of 15-LOX and caspase-3 as well as by transfection of PPAR-DN plasmid into A549 cells. A549 cells were treated with IL-4 (670 pM) and exposed to 5 M 15-PGJ2 and 30 M 15(S)-HETE for 72 h. The assay was performed with a cell death detection ELISA kit. Values represent the mean SD absorbance at 405 nm. Ap- optosis was also induced by incubation of cells with 5 M 15-PGJ2 and 30 M 15(S)-HETE. For blocking of 15(S)-HETE-induced apoptosis, cells were transiently transfected with PPAR-DN plasmid or were preincu- bated with 10 M NDGA 1 h before challenge with agonists. For inhibition of caspase-3, A549 cells were preincubated with 100 M Z-VAD-FMK, a caspase-3 inhibitor, for 1 h before adding IL-4. All experiments were per- formed in triplicate and represent the mean SD. B, IL-4 induces apo- ptosis in normal human bronchial epithelial cells. BEAS-2B cells were exposed to IL-4, 15(S)-HETE, and 15-PGJ2 as described above for exper- iments with A549 cells. Apoptosis was measured using a cell death detec- tion ELISA. The mean SD absorbance values at 405 nm are reported. , Significance compared with untreated cells (p 0.05).

Article Snippet: Nordihydroguaiaretic acid (NDGA; (Sigma-Aldrich, Deisenhofen, Germany) was used as a lipoxygenase inhibitor at a concentration of 10 M. Normal human bronchial epithelial cells (BEAS-2B) were obtained from American Type Culture Collection (Wesel, Germany) and were cultivated in a modified LHC-9 medium with all the necessary growth factors (supplied by Clonetics, Palo Alto, CA).

Techniques: Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Incubation, Blocking Assay, Inhibition

Journal: PloS one

Article Title: Infection of differentiated porcine airway epithelial cells by influenza virus: differential susceptibility to infection by porcine and avian viruses.

doi: 10.1371/journal.pone.0028429

Figure Lengend Snippet: Figure 3. Expression of sialic acid in swine PCLS cells. Sialic acids were detected by lectin staining: MAA (Maackia amurensis agglutinin) for a2,3-linked sialic acids and SNA (Sambucus nigra agglutinin) for a2,6- linked sialic acids. Ciliated cells were stained using an anti-b-tubulin antibody and mucus-producing cells were stained using an anti muc5ac antibody. In panels A, lectin-staining is compared with staining of ciliated (red) or mucus-producing cells (red in Ab, green in Ad); SNA staining is shown in green (Aa and Ab), MAA staining is shown in green (Ac) or red (Ad). In Ba, co-staining of ciliated (red) and mucus-producing cells (green) is shown; Bb shows co-staining with MAA (red) and SNA (green). In C samples were co-stained SNA (green) and MAA (red); cryosections were derived from PCLS before (Ca) or after neuraminidase treatment (Cb). doi:10.1371/journal.pone.0028429.g003

Article Snippet: To detect a2,6-linked sialic acids, FITC labeled Sambucus nigra agglutinin (SNA) (Vector laboratories, Burlungame, USA) was used and biotinylated Maackia amurensis lectin II (MAAII) was used to determine a2,3-linked sialic acids after preincubation of sections with the Avidin/Biotin Blocking kit (both from Vector Laboratories, USA).

Techniques: Expressing, Staining, Derivative Assay