Journal: Anesthesia & Analgesia

Article Title: The Pyrrole Etomidate Analog Carboetomidate Potently Inhibits Human 5-HT3A Receptor Function

doi: 10.1213/ane.0b013e31827d204d

Figure Lengend Snippet: Figure 1. Expression of 5-hydroxytryptamine type 3A (5-HT3A) protein in the human embryonic kidney (HEK)293 inducible cell line. No sig- nal was observed in uninduced HEK293 cells (control lane). Anti–5- HT3A immunoreactive protein from tetracycline-induced HEK293 cells expressing 5-HT3A cDNA has a molecular weight of approximately 50 kDa, similar to its predicted molecular weight. 5-HT3A protein aggre- gates were also observed at approximately 100, 200, and 250 kDa.

Article Snippet: Blots were then incubated with 1 μg/mL anti–5-HT3A antibody (goat polyclonal immunoglobulin G; Tocris Bioscience, Minneapolis, MN) in the blotting buffer overnight at 4°C.

Techniques: Expressing, Control, Molecular Weight

Journal: Anesthesia & Analgesia

Article Title: The Pyrrole Etomidate Analog Carboetomidate Potently Inhibits Human 5-HT3A Receptor Function

doi: 10.1213/ane.0b013e31827d204d

Figure Lengend Snippet: Figure 2. Carboetomidate and etomidate reversibly inhibit 5-hydroxytryptamine type 3A (5-HT3A) currents. A, Representative current traces of 5-HT3A currents elicited by 30-second application of 1 ìM 5-HT alone (left panel) and 10-second coapplication of 10 mM carboetomidate (middle panel) or etomidate (right panel) at midpoint during a 30-second application of 1 ìM 5-HT pulse. B, Percent inhibition (left panel) and recovery (right panel) of peak current amplitudes after application of carboetomidate or etomidate on 1 mM 5-HT elicited currents. Means ± SEM of n = 3 to 5, *** P < 0.0001, unpaired Student t test.

Article Snippet: Blots were then incubated with 1 μg/mL anti–5-HT3A antibody (goat polyclonal immunoglobulin G; Tocris Bioscience, Minneapolis, MN) in the blotting buffer overnight at 4°C.

Techniques: Inhibition

Journal: Anesthesia & Analgesia

Article Title: The Pyrrole Etomidate Analog Carboetomidate Potently Inhibits Human 5-HT3A Receptor Function

doi: 10.1213/ane.0b013e31827d204d

Figure Lengend Snippet: Figure 3. Concentration-response curves for inhibition of 5-hydroxytryptamine type 3A (5-HT3A) currents by carboetomidate and etomidate. A, Representative normalized current traces of inhibition of 5-HT3A currents by different concentrations of carboetomidate (left) and etomidate (right). 5-HT3A currents elicited by 100 μM 5-HT. B, Normalized integrated current (left panel), normalized peak amplitudes (middle panel), and normalized time constants of desensitization (right panel) as a function of hypnotic concentration. The control time constants of desensitiza- tion were 8.7 ± 0.8 seconds and 10.8 ± 0.8 seconds in the etomidate and carboetomidate groups, respectively. Data fit to Hill equation. Means ± SEM for 3 to 8 cells for each hypnotic concentration tested. IC50 = 50% inhibitory concentration.

Article Snippet: Blots were then incubated with 1 μg/mL anti–5-HT3A antibody (goat polyclonal immunoglobulin G; Tocris Bioscience, Minneapolis, MN) in the blotting buffer overnight at 4°C.

Techniques: Concentration Assay, Inhibition, Control

Journal: Anesthesia & Analgesia

Article Title: The Pyrrole Etomidate Analog Carboetomidate Potently Inhibits Human 5-HT3A Receptor Function

doi: 10.1213/ane.0b013e31827d204d

Figure Lengend Snippet: Figure 4. Carboetomidate enhances the 5-hydroxytryptamine type 3A (5-HT3A) current deactivation rates in outside-out patches. A, Representative normalized traces of 5-HT3A currents elicited by a 4-millisecond pulse of 100 μM 5-HT alone, or with 10 μM carboetomidate during deactivation. B, Mean ± SEM of deactivation time constants for control (closed squares, n = 7) and in the pres- ence of carboetomidate (closed circles; n = 5). * P < 0.043; unpaired Student t test.

Article Snippet: Blots were then incubated with 1 μg/mL anti–5-HT3A antibody (goat polyclonal immunoglobulin G; Tocris Bioscience, Minneapolis, MN) in the blotting buffer overnight at 4°C.

Techniques: Control

Journal: eLife

Article Title: Hsp47 promotes biogenesis of multi-subunit neuroreceptors in the endoplasmic reticulum

doi: 10.7554/eLife.84798

Figure Lengend Snippet:

Article Snippet: The pCMV6 plasmids containing human GABA A receptor α1 subunit ( GABRA1 ) (Uniprot #: P14867-1) (catalog #: RC205390), β2 subunit ( GABRB2 ) (isoform 2, Uniprot #: P47870-1) (catalog #: RC216424), γ2 subunit ( GABRG2 ) (isoform 2, Uniprot #: P18507-2) (catalog #: RC209260), 5-HT 3 A subunit (catalog #: SC122578, HTR3A , NM_000869), C-terminal FLAG-tagged 5-HT 3 B subunit (catalog #: MR206966, HTR3B , NM_020274), C-terminal FLAG-tagged hERG potassium channel (catalog #: RC215928, KCNH2 , NM_000238, human), and pCMV6 Entry Vector plasmid (pCMV6-EV) (catalog #: PS100001) were obtained from Origene (Rockville, MD, USA).

Techniques: Transfection, Construct, Control, Recombinant, Plasmid Preparation, Clone Assay, Software

Journal: Nature Communications

Article Title: Cryo-EM structure of 5-HT 3A receptor in its resting conformation

doi: 10.1038/s41467-018-02997-4

Figure Lengend Snippet: Data collection and processing

Article Snippet: The gene encoding 5-HT 3A R (purchased from GenScript USA Inc.) was inserted into a Xenopus laevis oocyte expression vector (pTLN) and confirmed by DNA sequencing.

Techniques: Microscopy, Glycoproteomics