ltl unconjugated  (Vector Laboratories)

Journal: Clinical and Experimental Nephrology

Article Title: Nephrotic syndrome induces the upregulation of cell proliferation-related genes in tubular cells in mice

doi: 10.1007/s10157-024-02608-1

Figure Lengend Snippet: Mki67 and Foxm1 were upregulated in various tubular cells. a Ki-67 immunostaining performed on 4 μm paraffin kidney sections. The number of Ki-67-positive cells was assessed in a minimum of 20 randomly selected high-power fields per animal (n = 3). **P < 0.01 (unpaired t-test). b Volcano plot of genes that have the ontology term of DNA-binding transcription factor activity (GO:0003700). c mRNA expression of Foxm1 and its downstream targets analyzed by the quantitative reverse-transcription polymerase chain reaction (n = 6). ***P < 0.001 (unpaired t-test). d Multiple in situ hybridization (ISH) procedures performed for Foxm1 and Mki67 . The sections were subjected to immunostaining of tubular markers after ISH. MKi67 -positive cells and Foxm1 -positive cells were counted in a minimum of 20 randomly selected high-power fields per animal (n = 3). **P < 0.01 and ****P < 0.0001 (unpaired t-test). Lotus tetragonolobus lectin (LTL), NKCC2, NCC, and AQP2 were utilized as markers for the proximal tubule, thick ascending limb, distal tubule, and collecting duct, respectively

Article Snippet: After the RNAscope protocol, the sections were subjected to heat-mediated antigen retrieval and stained with a rabbit polyclonal antibody specific to NKCC2 (catalog no.: ab191315, Abcam; diluted 1:400 in PBS), rabbit polyclonal antibody specific to NCC (catalog no.: AB3553, Sigma-Aldrich; diluted 1:400 in PBS), rabbit monoclonal antibody specific to AQP2 (catalog no.: ab199975, Abcam; diluted 1:1000 in PBS), and FITC-conjugated Lotus tetragonolobus lectin (Vector Laboratories) to identify the type of tubules.

Techniques: Immunostaining, Binding Assay, Activity Assay, Expressing, Reverse Transcription, Polymerase Chain Reaction, In Situ Hybridization

Journal: Clinical and Experimental Nephrology

Article Title: Nephrotic syndrome induces the upregulation of cell proliferation-related genes in tubular cells in mice

doi: 10.1007/s10157-024-02608-1

Figure Lengend Snippet: Mki67 and Foxm1 were upregulated in the adriamycin-induced nephrotic syndrome model. a Urine albumin, serum albumin, total cholesterol, and blood urea nitrogen (BUN) levels 14 days after adriamycin injection (n = 8). ***P < 0.001 (unpaired t-test). b mRNA expression of Mki67 and Foxm1 analyzed by quantitative reverse-transcription polymerase chain reaction (n = 8). c Multiple in situ hybridization (ISH) procedures performed for Foxm1 and Mki67 . The sections were subjected to immunostaining for tubular markers after ISH. ***P < 0.001 (unpaired t-test). Lotus tetragonolobus lectin (LTL), NKCC2, NCC, and AQP2 were utilized as markers for the proximal tubule, thick ascending limb, distal tubule, and collecting duct, respectively. d The number of Mki67 -positive cells and Foxm1 -positive cells was counted in a minimum of 20 randomly selected high-power fields per animal (n = 3). **P < 0.01 (unpaired t-test)

Article Snippet: After the RNAscope protocol, the sections were subjected to heat-mediated antigen retrieval and stained with a rabbit polyclonal antibody specific to NKCC2 (catalog no.: ab191315, Abcam; diluted 1:400 in PBS), rabbit polyclonal antibody specific to NCC (catalog no.: AB3553, Sigma-Aldrich; diluted 1:400 in PBS), rabbit monoclonal antibody specific to AQP2 (catalog no.: ab199975, Abcam; diluted 1:1000 in PBS), and FITC-conjugated Lotus tetragonolobus lectin (Vector Laboratories) to identify the type of tubules.

Techniques: Injection, Expressing, Reverse Transcription, Polymerase Chain Reaction, In Situ Hybridization, Immunostaining