Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: Targeting the Gdnf Gene in peritubular myoid cells disrupts undifferentiated spermatogonial cell development
doi: 10.1073/pnas.1517994113
Figure Lengend Snippet: Antibodies
Article Snippet: Species Dilution Primary ACTA2 Sigma-Aldrich, A5228 (Clone 1A4) Mouse, monoclonal 1:200 ACTA2-FITC Sigma-Aldrich, F3777 (Clone 1A4) Mouse, monoclonal 1:200 dtTomato Clontech, 632496 Rabbit, polyclonal 1:500 GATA4 Santa Cruz, SC-1237 Goat, polyclonal 1:300 GDNF ( d -20) Santa Cruz, SC-328 Rabbit, polyclonal 1:200 GFP Life Technologies, {"type":"entrez-nucleotide","attrs":{"text":"A11122","term_id":"490966","term_text":"A11122"}} A11122 Rabbit, polyclonal 1:200 HSPA2 Eddy laboratory, Allen et al. PMID 3352605 Rabbit, polyclonal 1:1,000 KI67 Abcam, ab15580 Rabbit, polyclonal 1:200 KIT Abcam, ab65525 (Clone 2B8) Rat, monoclonal 1:100 SOX9 Cosmo Bio Co. Japan, KAL-KO608 Rabbit, polyclonal 1:200 ZBTB16 (PLZF, H-300) Santa Cruz, SC-22839 Rabbit, polyclonal 1:200 Secondary Alexa Fluor 488, Donkey anti-mouse IgG Life Technologies, {"type":"entrez-nucleotide","attrs":{"text":"A21202","term_id":"641355","term_text":"A21202"}} A21202 Donkey, polyclonal 1:200 Alexa Fluor 546, Donkey anti-rabbit IgG Life Technologies, {"type":"entrez-nucleotide","attrs":{"text":"A10040","term_id":"489103"}} A10040 Donkey, polyclonal 1:200 Alexa Fluro 647, Donkey anti-goat IgG Life Technologies, {"type":"entrez-nucleotide","attrs":{"text":"A11056","term_id":"489255","term_text":"A11056"}} A11056 Donkey, polyclonal 1:200 DAB staining VECTSTAIN Elite ABC Kit (Rabbit IgG) Vector Laboratories, PK-6106 ImmPACT DAB Peroxidase Substrate Vector Laboratories, SK-4105 Vybrant apoptosis Assay Kit 6 (V23200) Annexin V + Alexa Fluor 350 Life Technology PI Life Technology Open in a separate window Antibodies fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. S2. caption a7 ZBTB16 immunostaining of undifferentiated spermatogonia in testes from WT, Het, and cKO mice at 1, 2, 4, 8, and 12 wk. ( A ) No obvious differences are seen at 1 wk. ( B ) Fewer ZBTB16-positive cells were present in cKO testes compared to Het and WT at 2 wk. ( C – E ) ZBTB16-positive cells were rarely observed at 4 ( C ), 8 ( D ), and 12 ( E ) wk in cKO testes.
Techniques: Staining, Plasmid Preparation, Apoptosis Assay