Journal: Developmental biology
Article Title: Epithelial DNA methyltransferase-1 regulates cell survival, growth and maturation in developing prostatic buds
doi: 10.1016/j.ydbio.2019.01.011
Figure Lengend Snippet: (A) RT-PCR for Cdkn1a mRNA in E18.5 control and cDnmt1KO urethras. (B-C) E18.5 urethra sections were labeled with antibodies against Gamma-H2AX (in red, DNA damage marker) and LacZ (in green, labels Shh lineage epithelium). (D-E) E18.5 urethra sections were labeled with antibodies against Phospho-p53 Ser15 (in red, marks active p53). (F-G) E18.5 urethra sections were labeled with antibodies against Cleaved caspase 3 (in red, marks apoptotic cells) and EYFP (in green, labels Shh lineage epithelium). Green channel is excluded for ease of visualization. (H) Percentage of cleaved Caspase 3 labeled cells in the Shh lineage urethral epithelium. White arrowheads indicated cleaved Caspase 3 positive apoptotic cells. White dotted lines indicate epithelial-mesenchymal interface. DAPI staining is shown in blue. Scale bar is 50 microns. Graphical results are the mean + SEM of at least three mice per group. p-values indicate significant differences (* p<0.05, ** p<0.01) between groups based on Student’s unpaired t-test.
Article Snippet: For primary and secondary antibody information see . table ft1 table-wrap mode="anchored" t5 Table 1 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-5 methylcytosine Abeam Cat#ab10805; RRID:AB_442823 Chicken polyclonal anti-LacZ Abeam Cat#ab9361, RRID:AB_307210 Chicken polyclonal anti-GFP Abeam Cat#ab13970; RRID:AB_300798 Rabbit polyclonal anti-Ki67 Abeam Cat#ab15580; RRID:AB_443209 Rabbit polyclonal anti-Gamma Tubulin Abeam Cat#ab11317; RRID:AB_297921 Rabbit polyclonal anti-PAX2 Covance Cat#PRB-276P-200; RRID:AB_291611 Mouse monoclonal anti-FOXA1 Millipore Cat#05–1466; RRID:AB_1977191 Chicken polyclonal anti-KRT5 Biolegend Cat#905901; RRID: AB 2565054 Rabbit monoclonal anti-5 methylcytosine Cell Signaling Cat#28692 Rabbit monoclonal anti-DNMT1 Cell Signaling Cat#D63A6; RRID:AB_10828695 Rabbit monoclonal anti-CDH1 Cell Signaling Cat#3195S Rabbit polyclonal anti-Histone H2AX (phospho Ser139) Cell Signaling Cat#2577; RRID:AB_2118011 Mouse monoclonal anti-p53 (phospho Ser15) 16G8 Cell Signaling Cat#9286; RRID:AB_331741 Rabbit monoclonal anti-Cleaved Casp3 (Asp 175) Cell Signaling Cat#9664; RRID:AB_2070042 Rabbit polyclonal anti-Phospho-Histone H3 (Ser10) Cell Signaling Cat#9701; RRID:AB_331535 Mouse monoclonal anti-CDH1 BD Transduction Labs Cat#610181; RRID:AB_397580 Mouse monoclonal anti-ZO1 Thermo Fisher Scientific Cat#33–9100; RRID:AB_2533147 Mouse monoclonal anti-KRT14 Thermo Fisher Scientific Cat#MS-115-P0, RRID:AB_63786 Goat polyclonal anti-Chicken Alexa488 conjugated Jackson Immunoresearch Cat#103–545–155; RRID:AB_2337390 Goat polyclonal anti-Mouse Alexa488 conjugated Jackson Immunoresearch Cat# 115–547–003; RRID:AB_2338869 Goat polyclonal anti-Rabbit Alexa488 conjugated Jackson Immunoresearch Cat# 111–547–003; RRID:AB_2338058 Goat polyclonal anti-Mouse Alexa594 conjugated Jackson Immunoresearch Cat# 115–585–062; RRID:AB_2338876 Goat polyclonal anti-Rabbit Alexa594 conjugated Jackson Immunoresearch Cat# 111–586–045; RRID:AB_2338067 Sheep polyclonal anti-Digoxygenin Alkaline phosphatase conjugated Roche Cat#11093274910; RRID:AB_514497 Chemicals, Peptides, and Recombinant Proteins Tamoxifen, free base Sigma Aldrich T5648;CAS: 10540–29–1 5-ethynyl-2’-deoxyuridine (EdU) Thermo Fisher A10044 Critical Commercial Assays Click-iT® EdU Alexa Fluor® 488 Imaging Kit Thermo Fisher C10337 Deposited Data RNA-Seq data This paper GEO: GSE121086 Experimental Models: Organisms/Strains Mouse: B6.129S4- Dnmt1 tm2Jae/Mmucd MMRRC, UC Davis Stock#014114-UCD; RRID:MMRRC_0141 14-UCD Mouse: B6.
Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Labeling, Marker, Staining
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