Dsc hx300

A JL-A electronic stimulator (Shanghai, China) was used to perform the electrical stimulation. A unidirectional square pulse of constant pulse width (0.6 ms) with no interphase gap, 40 Hz in stimulation frequencies and 50 V in strength were applied for 0.25 s. The electrode was placed on a portion of the excised skin pieces and the rest of the skin was used as a control. Images were taken using a SONY camera (DSC-HX300) before and after the electrical stimulation and 1 min after the cessation of stimulation. The experiment was repeated three times on three different fish.

Forskolin (Selleck Chemicals, Houston, TX, USA), an AC stimulating agent, was first dissolved in DMSO (Solarbio, Beijing, China) at a concentration of 10 mM, and then diluted in PBS to a final concentration of 100 μM, which was applied to the blue spots of the body surface, and the color change of spots was recorded with SONY camera (DSC-HX300). The experiment was repeated three times on three different fish. The changes in diameter of spots in the color change process were assessed by Nano Measurer 1.2 (n=30 spots). We used online tools Image Color Detection to extract the hue of the blue (n=6 spots) and black spots (n=6 spots) and obtain the color code. Then, we obtained the corresponding HSL and RGB values with the help of the color code converter in Rappidtables.

4-week-old female BALB/c nude mice were purchased from Shanghai Lingchang Biotechnology Co., Ltd. to construct a tumor nude mouse model by subcutaneous injection, and 10 nude mice were randomly assigned to each group. All animal experiments were approved by the Ethics Committee of Laboratory Animal Care and Use of Peking University First Hospital (Approval No. J2020-14). All animals were treated according to the standards prescribed by the “Guide-lines for the welfare and use of animals in cancer research”. MGC-803 cells (4.0 ×10⁶ cells) infected with shPSMC2 and negative control shRNA lentivirus were injected subcutaneously into the right forelimb armpit of nude mice. The tumor volume and mouse weight were measured 13 days after injection to draw the tumor growth curve. After 22 days of injection, nude mice were anesthetized by intraperitoneal injection of 0.7% Pentobarbital Sodium at 10 μl/g dose. Then nude mice were placed on an in vivo imaging instrument (LB983, Berthold Techologies) for imaging, and the fluorescence intensity was observed. After the nude mice were sacrificed, the tumors were taken out from the nude mice and photographed with a digital camera (DSC-HX300, SONY). The tumor volume and weight were measured. Immunohistochemical staining was performed in accordance with the method described above to evaluate the expression of Ki67 in tumor tissues.

AGS and MGC-803 cells infected with shRNA or shPSMC2 lentivirus were transferred to 6-well plate with 500 cells per well, and cultured for 8 days. The cells were washed once with PBS, and then 1 ml of 4% polyformaldehyde (Sinopharm Chemical Reagent Co., Ltd) was added to fix the cells for 30–60 min. The cells were washed once with PBS, and the GIEMSA dye (AR-0752, Shanghai Dingguo Biotechnology Co. Ltd) was added to each well for staining for 15 min. Cells were washed with ddH₂O, and taken pictures with a with digital cameras (DSC-HX300, SONY) after drying. The number of clones was counted (each clone contains more than 50 cells). There were 3 independent repetitions in this assay.