Journal: Cell reports

Article Title: A High-Content Screen Identifies TPP1 and Aurora B as Regulators of Axonal Mitochondrial Transport

doi: 10.1016/j.celrep.2019.08.035

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: pCMV6-Myc-DDK-Aurkb (human) , OriGene , RC210288.

Techniques: Recombinant, Software

Journal: The Journal of Neuroscience

Article Title: A Syntaxin 1, Gα o , and N-Type Calcium Channel Complex at a Presynaptic Nerve Terminal: Analysis by Quantitative Immunocolocalization

doi: 10.1523/JNEUROSCI.0346-04.2004

Figure Lengend Snippet: Intensity correlation analysis, experimental models. Each panel in A, C, and E shows a calyx nerve terminal stained with two antibodies with the overlay on the left and the individual B/W image pairs on the right. Each panel in B, D, and F shows intensity correlation plots on the left together with the respective plots of stain A (center) and B (right) pixel intensities against their (A - a)(B - b) values (see Results, Intensity correlation analysis). A, B, Experimental example of dependent staining: calyx nerve terminal stained for SV2 with monoclonal (red, left panel; B/W center panel) and polyclonal (green, left panel; B/W right panel) tagged antibodies. ICQ = 0.48, psign test < 0.001. C,D, Experimental example of segregated staining. Calyx terminal stained for SV2 (red, left panel; B/W center panel) and Na/K pump (green, left panel; B/W right panel). ICQ = -0.19, psign test < 0.001. E, F, Experimental example of complex staining. A calyx nerve terminal stained with a mAb against SV2 (red, left panel; B/W center panel) and polyclonal antibody against neurofilament (green, left panel; B/W right panel). Scale bars, 5 μm.

Article Snippet: Affinity-purified polyclonal antibodies were: neurofilaments, AB1991 (Chemicon, Temecula, CA); MUNC18 (ABR); syntaxin (Sigma); SV2 (Stressgen, Vancouver, British Columbia, Canada); Ca V 2.1, CW65 ( Fletcher et al., 2001 ), Ca V 2.2, CW21 (also termed CW14; McEnery et al., 1997 ).

Techniques: Staining

Journal: The Journal of Neuroscience

Article Title: A Syntaxin 1, Gα o , and N-Type Calcium Channel Complex at a Presynaptic Nerve Terminal: Analysis by Quantitative Immunocolocalization

doi: 10.1523/JNEUROSCI.0346-04.2004

Figure Lengend Snippet: Ab571 staining localized release sites at the chick ciliary ganglion calyx nerve terminal. A, Primary hippocampal neuron culture stained with Ab571 (green) and monoclonal anti-syntaxin (red). Note Ca channel puncta are along the same trajectories as the syntaxin staining, consistent with the synaptic contacts formed by a nerve fiber process onto an underlying postsynaptic neuron (data not shown). B, As for A except double-stained with monoclonal anti-SV2 (red). Some colocalization of the two dyes was noted in both A and B, but puncta of single stained syntaxin, SV2, or Ca channels were also evident. C, An optical section through a calyx presynaptic terminal attached to postsynaptic ciliary neuron stained for Ca channels (Ab571; green, top panel) and both Ab571 and synaptic vesicle clusters (SV2; red, bottom panel). A diagram (not to scale) of the synapse is presented in the bottom panel. D, Calyx terminal stained with Ab571 (top left panel) rabbit polyclonal SV2 (top center panel), and Na-K pump (top right panel) using the pretty-poly multiple polyclonal immunostaining technique (see Results). The next three (middle row) panels show colocalization of Ab517 and SV2 (left panel, middle row), Ab571 and Na-K pump (center panel, middle row), and SV2 and Na-K pump (right panel, middle row). The bottom panel is a triple overlay of the three stains. The ciliary neuron is above the calyx. Note the Ca channel clusters on the transmitter release face aspect and the secretory vesicle clouds overlapping the channels and distal from the release site. The staining in the image has been adjusted digitally to enhance the resolution of the three structures. Some distortion of sizes (such as surface membrane thickness) is a result of both the membrane coursing through the image plane at an angle and of limitations in the resolution of the light microscope. Scale bars: A, B, 5 μm; C,D, 2 μm.

Article Snippet: Affinity-purified polyclonal antibodies were: neurofilaments, AB1991 (Chemicon, Temecula, CA); MUNC18 (ABR); syntaxin (Sigma); SV2 (Stressgen, Vancouver, British Columbia, Canada); Ca V 2.1, CW65 ( Fletcher et al., 2001 ), Ca V 2.2, CW21 (also termed CW14; McEnery et al., 1997 ).

Techniques: Staining, Immunostaining, Light Microscopy