Journal: The Journal of Neuroscience

Article Title: α 2 δ-4 Is Required for the Molecular and Structural Organization of Rod and Cone Photoreceptor Synapses

doi: 10.1523/JNEUROSCI.3818-16.2018

Figure Lengend Snippet: Antibodies used for immunofluorescence or Western blot (WB)

Article Snippet: The working concentrations of these and other antibodies are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Source Catalog Concentration RRID Rabbit anti-α 2 δ-4 In house (this paper) NA 1:1000 NA (current study) Rabbit anti-Ca v 1.4 In house ( Liu et al., 2013 ) NA 1:1000–1:4000 AB_2650487 Rabbit anti-Ribeye Synaptic Systems 192103 1:1000 AB_2086775 Mouse anti-Ribeye BD bioscience 612044 1:1000 1:500 (WB) AB_399431 Mouse anti-PSD95 Thermo Fisher Scientific MA1-046 1:1000 AB_2092361 Mouse anti-PSD-95 Millipore 529541 1:1000 (WB) AB_565012 Rabbit anti-ERC 1b/2 (CAST) Synaptic Systems 143003 1:2000 (WB) AB_887715 Rabbit anti-synaptobrevin2 (VAMP2) Synaptic Systems 104202 1:5000 (WB) AB_887810 Rabbit anti-PKC-α Santa Cruz Biotechnology SC208 1:500 AB_2168668 PNA-AF647 Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"L32460","term_id":"497530","term_text":"L32460"}} L32460 1:250 NA Sheep anti-mGluR6 K. Martemyanov NA 1:200 AB_2650490 Guinea pig anti-mGluR6 Neuromics GP13105 1:500 AB_2341540 Mouse anti-GluR2 Millipore MAB397 1:150 AB_11212990 Mouse anti-calbindin Sigma-Aldrich C9848-100UL 1:400 AB_476894 Rabbit anti-secretagonin Biovendor RD181120100 1:1000 AB_2034060 Mouse anti-HA Covance MMS-101R 1:1000 (WB) AB_291263 Mouse anti-β-actin Sigma-Aldrich A2228 1:10 000 (WB) AB_476697 AF488 goat anti-mouse Thermo Fisher Scientific A-11001 1:500 AB_2534069 AF568 goat anti-mouse Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"A11004","term_id":"492388","term_text":"A11004"}} A11004 1:500 AB_2534072 AF647 goat anti-mouse Thermo Fisher Scientific A-21235 1:500 AB_2535804 AF488 goat anti-rabbit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"A11008","term_id":"492390","term_text":"A11008"}} A11008 1:500 AB_143165 AF568 goat anti-rabbit Thermo Fisher Scientific A-11011 1:500 AB_143157 AF647 goat anti-rabbit Thermo Fisher Scientific A-21244 1:500 AB_2535812 AF488 donkey anti-sheep Thermo Fisher Scientific A-11015 1:500 AB_2534082 AF647 donkey anti-sheep Thermo Fisher Scientific A-21448 1:500 AB_2535865 AF488 donkey anti-guinea pig Thermo Fisher Scientific A-11073 1:500 AB_2534117 Streptav-AF488 Thermo Fisher Scientific {"type":"entrez-protein","attrs":{"text":"S11223","term_id":"112468","term_text":"pir||S11223"}} S11223 1:500 AB_2336881 Biotin conjugate Thermo Fisher Scientific 31820 1:500 AB_228340 HRP sheep anti-mouse IgG GE Healthcare NA931 1:20,000 (WB) AB_772210 HRP donkey anti-rabbit IgG GE Healthcare NA934V 1:20,000 (WB) AB_772206 AF594-WGA Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"W11262","term_id":"1285567","term_text":"W11262"}} W11262 1:1000 NA Open in a separate window Antibodies used for immunofluorescence or Western blot (WB)

Techniques: Immunofluorescence, Western Blot, Concentration Assay

Journal: Cell death discovery

Article Title: Inhibition of Aurora Kinase B activity disrupts development and differentiation of salivary glands.

doi: 10.1038/s41420-020-00393-w

Figure Lengend Snippet: Fig. 8 AURKB loss disrupted acinar identity via ROS generation. A, B Mist1 (green) and pan cytokeratin (blue) expressions in E16.5+24h bud cells, scale bar = 50 µm. Remarkable loss of the acinar differentiation marker Mist1 at the protein and transcriptional levels in response to Barasertib treatment. Recovery of Mist1 levels due to ROS scavenging using aminoguanidine (number of pooled glands ≥5), analysed by ANOVA **P ≤0.01, *p ≤0.05, ****P ≤0.0001. C, D ROS production in response to AURKB inhibition in the embryonic salivary glands. ROS levels were visualised in the SMG explants after staining with DCFDA and inspection under fluorescence microscope. Analysis of corrected total fluorescence (CTF) using ImageJ revealed that Barasertib treatment for 24 h significantly increased levels of ROS in the E16.5+24h explants which were significantly reduced after co- treating the explants with aminoguanidine, analysed by ANOVA, **P ≤0.01, *p ≤0.05, ns: non-significant, p > 0.05. Scale bar = 500 μm, n = 3/group.

Article Snippet: To block endogenous peroxidase activity and nonspecific background staining sections were incubated in 3% hydrogen peroxide solution for 20–30 min. To block all epitopes on the tissue samples and prevent nonspecific antibody binding, sections were incubated with 1% BSA in 1X TBS, pH 7.6 for 5 min. Tissue sections were incubated at 4 °C overnight, with the antibody: rabbit-anti-cleaved caspase 3 (1:2500, NB100–56113, Novus Bio), rabbit–anti-Mist1 (1:200, ab187978, Abcam), mouse-anti-Cyclin D1 (1:400, NBP2–32840, Novus Bio), rabbit-anti-AURKB (1:200, ab115793, Abcam), rabbit-anti-ki67 (1:100, ab16667, Abcam), rabbit-anti-p21 (1:200, ab188224, Abcam), rabbit-anti-p.H2AX(1:200, ab81299, Abcam), rat-anti-E- Cadherin (1:200, sc-59778, Santa Cruz Biotechnology, Inc.) guinea pig-anti-pan cytokeratin (1:50, BP5069, Origene).

Techniques: Marker, Inhibition, Staining, Microscopy