After 1-week acclimation, mice were randomly assigned into four groups (n = 6): Control group, ITP group, ITP + vector group and ITP + sh-HLA-DRB5 group. Before establishment of ITP murine models, guinea pig anti-mouse platelet serum (GP-APS) that can be used to induce ITP was customized and purchased from Cloud-Clone Corp. (Wuhan, China). Initially, mice received injection of serum containing GP-APS, and then PLT was removed from the circulation by the fixed phagocyte system [18 (link)]. In this study, GP-APS was diluted with saline (G4702, Servicebio, Wuhan, China) in a ratio of 1:4 to prevent erythrocyte adsorption [19 (link)]. Then, besides mice from Control group that were given the same volume of saline, mice from other groups were intraperitoneally injected with 100 μL GP-APS every other day. Meanwhile, mice from ITP + sh-HLA-DRB5 group or ITP + vector group were injected with 50 μL diluted sh-HLA-DRB5 adenovirus or negative control by tail vein once a week for 2 weeks. After 15 days, all mice were euthanized by cervical disarticulation under anesthesia (50 mg/kg pentobarbital sodium, P-010; Sigma-Aldrich, St Louis, MO, USA).
P 010
Manufactured by Merck Group
Sourced in United States, Germany, China
The P-010 is a versatile laboratory equipment that serves as a high-performance centrifuge. It is designed to efficiently separate and isolate a variety of samples, including cells, particles, and macromolecules, through the application of centrifugal force. The core function of the P-010 is to facilitate the separation and purification processes that are essential in various scientific and research applications.
Automatically generated - may contain errors
Lab products found in correlation
G4702, by Wuhan Servicebio Technology (1 mentions)
Balb c nude mice, by Charles River Laboratories (1 mentions)
Mab360, by Merck Group (1 mentions)
Lps l2630, by Merck Group (1 mentions)
Bh 2 microscope, by Olympus (1 mentions)
P5669, by Merck Group (1 mentions)
Cerulein, by AnaSpec (1 mentions)
Male c57bl 6 mice, by Shanghai Model Organisms (1 mentions)
D8418, by Merck Group (1 mentions)
Sprague dawley rat, by Charles River Laboratories (1 mentions)
29 protocols using p 010
1
Induction of Immune Thrombocytopenia in Mice
2
Investigating FAM201A and miR-1271-5p in Xenograft Tumor Model
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BALB/c nude mice (Male, 5–6-week-old) were purchased from the Vital River Laboratories (Beijing, China). The mice were maintained under a specific condition (22~ 24°C, 50% humidity, a 12 h:12 h circadian cycle), with free access to a standard mice chow and water. Then, the mice were randomized into four groups (n = 6 per group): NC+MC group, FAM201A+MC group, NC+miR-1271-5p M group, and FAM201A+miR-1271-5p M group. After transfection, SiHa cells (5 × 106) with stable expressions of FAM201A, miR-1271-5p or both were subcutaneously injected into the posterior flank of the mice. The size of subcutaneous xenografts (length and width) was measured by a caliper every 7 days, with 5 times in total, and the volume of the xenografts was calculated according to the formula: 0.5 × length × width2. Five weeks after the injection, the mice were sacrificed via spinal dislocation under anesthetization using pentobarbital sodium (P010, Sigma-Aldrich, USA), following which the subcutaneous xenografts were resected and weighed.
3
Immunohistochemical Analysis of Mouse Brain
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Mice in different treatment groups were anesthetized with pentobarbital sodium (Sigma-Aldrich, P-010) and perfused with PBS followed by 4% paraformaldehyde in PBS. The brains were harvested and post-fixed in 4% paraformaldehyde, followed by dehydration in 20% sucrose-PBS and then 30% sucrose-PBS. Next, the brains were cut into frozen slices (25 µm). After being rinsed with PBS, the sections were treated with 3% H2O2 for 15 min to quench endogenous peroxidase activity, permeabilized and blocked with 0.3% Triton X-100 in PBS containing 5% BSA for 1.5 h at room temperature. Then, the slices were incubated with primary TH antibody (Sigma-Aldrich, T1299) and primary GFAP antibody (Millipore, MAB360) at 4 °C overnight and then with the corresponding secondary antibody, and the results were visualized by the DAB reaction. For Nissl staining, slices were stained in 0.1% cresyl violet acetate solution (0.1 g cresyl violet in 99% H2O and 1% acetic acid) for 30 min at room temperature. Then, slices were dehydrated with ethanol and xylene. Stereo Investigator software was used to image and count the number of positive cells under the microscope (Olympus BX51).
4
Spinal Cord Compression Injury in Mice
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The mice were anesthetized with 50 mg/kg pentobarbital sodium (P-010; Sigma, St. Louis, Missouri, United States) and underwent surgery in the prone position. The back hair was shaved, and the skin was disinfected with iodophor 3 times. All surgical instruments were autoclaved. A longitudinal incision in the skin was made at the level of T10 and extended down until the lamina was exposed. A rongeur was used to remove part of the lamina of T10 until the spinal cord was exposed. Fine forceps with a width of 0.5 mm at the tip (511,252–20, Fine Science Tools, Heidelberg, Germany) were used to apply pressure from both sides of the spinal cord for 5 seconds [14 ]. During the clamping process, the spinal cord was clamped completely and the same clamping pressure was maintained (with the tips of the fine forceps just touching as the standard), resulting in moderate compression injury. To reduce the influence of interfering factors on the functional recovery of mice, those with poor postoperative states or severe complications were excluded as experimental subjects. After the operation, the bladder was pressed to assist the mice in urinating twice a day. On the first day after surgery, D-4F was continuously injected intraperitoneally (dissolved in phosphate-buffered saline (PBS) at a dose of 1 mg/kg/day per day), while the control groups were injected with the same amount of PBS.
5
Spinal Cord Injury Mouse Model
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The mice were anesthetized by intraperitoneal injection of 2% pentobarbital sodium (P-010; Sigma, St. Louis, MO, USA) and placed in the prone position. The back was then shaved at the T10 level, and the skin was disinfected with iodophor. Bone forceps were used to remove the lamina to expose the spinal cord at the T10 level, and Dumont forceps with a 0.5-mm-wide tip (No. 5, 11252-20, Fine Science Tools, Heidelberg, Germany) were used to completely compress the spinal cord from both sides for 5 seconds, resulting in moderate compression injury (Wanner et al., 2013). After the SCI was induced, the bladder was pressed twice a day to assist mice in urinating until their urination reflex recovered. Before SCI induction, the mice were randomly divided into four groups: pre-injury (n = 8) and 3 (acute phase, n = 14), 7 (subacute phase, n = 14), and 14 (subacute phase, n = 14) dpi. The total number of animals used in this study was 50. Three mice died due to intraoperative blood loss.
6
LPS-Induced Acute Lung Injury Model
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Fifty-four 6- to 8-week-old male C57BL/6 mice (20∼25g) were purchased from ALF Biotechnology (Jiangsu, China). All the mice were maintained at 20∼24℃ and 50% humidity on a 12 h/12 h light/dark cycle, and were given free access to food and water. Prior to experimen-tation, the mice were acclimated for 3 days.
All the mice were anesthetized via intraperitoneal injection of 2% pentobarbital sodium (P-010, Sigma-Aldrich, USA). For establishment of ALI models, the mice were intratracheally injected with a single dose of 100 μg lipopolysaccharide (LPS, L2630, Sigma-Aldrich, USA) which was previously dissolved in 50 μl normal saline (NS). The mice were allowed to recover until fully awake. The lentivirus-infected BMSCs (5×104 cells) were resuspended in 30 μl of PBS for implantation therapy.
All the mice were anesthetized via intraperitoneal injection of 2% pentobarbital sodium (P-010, Sigma-Aldrich, USA). For establishment of ALI models, the mice were intratracheally injected with a single dose of 100 μg lipopolysaccharide (LPS, L2630, Sigma-Aldrich, USA) which was previously dissolved in 50 μl normal saline (NS). The mice were allowed to recover until fully awake. The lentivirus-infected BMSCs (5×104 cells) were resuspended in 30 μl of PBS for implantation therapy.
7
Histological Analysis of Vascular Structures
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The rats were anesthetized via intra-abdominal injection with pentobarbital (100 mg/kg, Sigma-Aldrich, P-010) and euthanized by cervical dissociation. The thoracic aortas and superior mesenteric arteries were fixed in 4% paraformaldehyde (Sigma-Aldrich, 158127) for 7 days, soaked in 30% sucrose, embedded in paraffin wax, sectioned (4 μm), and then stained with haematoxylin and eosin as described previously [19 (link)]. The sections were visualized under an Olympus BH-2 microscope. The values of vascular wall thickness, cross-sectional area, and vascular smooth muscle cells (VSMCs) width were quantified by using ZEN 2.3 (blue edition). The investigators were blinded for acquiring and quantifying the images.
8
Intraocular Pressure-Induced Retinal Ischemia Model
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The establishment of an I/R was applied according to our prior study.23 Specifically, intraperitoneal injection of 1% pentobarbital sodium (P-010; Sigma Chemical Co., St. Louis, MO, USA) was used for the anesthesia of mice (100 mg/kg). In addition, corneal anesthesia was achieved by applying 1 to 2 drops of 0.5% tetracaine (H20071092; Zhongshan Ophthalmic Center, Guangzhou, Guangdong, China) topically, and 1 to 2 drops of 0.5% tropicamide phenylephrine eye drops (H20055546; Zhongshan Ophthalmic Center, Guangzhou, Guangdong, China) were administered to ensure pupillary dilation. The IOP in the test eye was increased to 110 mm Hg for 1 hour by the perfusion of anterior chamber with a 32-gauge aseptic needle connected to a suspended saline sack at the height of 1.5 m. The IOP was measured using a handheld digital rebound tonometer (TonoLab; Icare, Espoo, Finland). Retinal ischemia was confirmed by whitening of the iris and loss of the red reflex.3 (link),24 (link) Then, the needle was withdrawn to allow for retinal reperfusion, which was evidenced by the recovery of the red reflex.24 (link) The fellow eye was injected with a 32-gauge aseptic needle but without elevation of the saline sack and served as a control group. Eyes that develop cataracts or infections theoretically should be excluded, even though none of the eyes in the current research actually developed either.
9
Corneal Nerve Severing in Mice
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All surgeries were performed on eight-week-old female mice under anesthesia with the aid of an operating microscope. Mice were anesthetized by intraperitoneal injection of 1% pentobarbital sodium (P-010; Sigma-Aldrich Corp., St. Louis, MO, USA). To mimic refractive surgery, a circular punch (Kai Europe GmbH, Solingen, Germany) was used to sever the stromal nerves of the right eye cornea. The punch was applied to the corneal surface and then twisted five times with slight pressure until the stroma was incised.16 (link) At the end of the surgery, a drop of tobramycin eyedrops was applied to the nerve severing eye. The right eyes and left eye of the corneal nerve-cutting group were defined as test (T) and contralateral (CL) eyes, respectively. The right eye was lightly pressed with a cotton swab in sham group.
10
Autistic Rat Models with VB6 Deficiency
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Autistic rat models with VB6 deficiency or VB6 supplement were constructed using the offspring of female Wistar rats as previously described [12 (link)]. Based on a basic diet, all female rats were randomly fed with 0, 6, or 30 mg VB6 (P5669, Sigma-Aldrich, St. Louis, MO, USA) for 4 weeks. During the pregnancy and lactation, in consistent with the grouping of their mother rats, the rats were continued to be given aforementioned VB6 treatments. After weaning, new pups (male) were collected and divided into the three groups: Control group (n = 6, a basic diet with 6 mg VB6), VB6 deficiency group (n = 24, a basic diet without VB6), and VB6 supplement group (n = 6, a basic diet with 30 mg VB6). The body weight of offspring rats was recorded from postnatal day 1 (PND 1) to PND 42. The offspring rats between PND 42 and PND 56 were subjected to the behavioral tests. The mother rats were sacrificed by cervical dislocation under anesthesia (40 mg/kg, pentobarbital sodium, P-010, Sigma-Aldrich, USA).
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