The male C57BL/6 mice (8 weeks old, 22-25 g body weight) used in this study were purchased from Dashuo Bio-Technique Co. Ltd. (Chengdu, China) and were maintained in 12 hr of light/12 hr of darkness. The mice were randomly divided into the following 4 groups (n = 8 per group): Sham, UUO, UUO with tectorigenin (20 mg/kg/day, dissolved in saline with 10% dimethyl sulfoxide [DMSO]) or irbesartan (IRB, a positive control, 20 mg/kg/day) treatment. After anesthesia with 1% pentobarbital, the left ureter was exposed through the lower left incision on the midline of the back and blocked by two-point ligation with 40 silk thread as previously described. The Sham-operated mice underwent the same operation but absence of ligation. The tectorigenin and IRB group were administered intraperitoneally with drug daily for 7 consecutive days since surgery. Sham-operated mice and UUO mice were received the equal volume of solvent.

The athymic BALB/c nude mice (4 weeks; 20-22 g; Beijing Vital River Laboratory Animal Technology Company, China) were housed in a specific pathogen-free environment under a 12-h lightdark cycle with free access to food and water. The animals were allowed to acclimatize to their surroundings for 3 days. U87 cells (1 x 10⁶) in the logarithmic growth phase in 100 uL PBS were subcutaneously injected into the right flank. Therapeutic experiments were started when the tumor reached around 150 mm³ after about 10 days. Mice were allocated to receive intraperitoneal injections of vehicle (control group, n = 6) or 40 mg/kg bodyweight (n = 6) in the same volume (50 uL) once a day for 13 times.

Present animal studies used male Sprague Dawley rats (80-100 g) to create a HF model induced by the descending aortic banding (AB) procedure. The sham-operated (SO) group was defined as rats subjected to a similar procedure with the exception of arterial ligation. Echocardiography was applied to confirm the arterial banding immediately after the surgical procedure.

Adult male C57BL6 (C57) mice (8-12 weeks, 20-25 g) were purchased from the Animal Experiment Center of Wuhan University. All 64 mice were randomly divided into various groups by different treatments (n = 8). In sham group, after the right kidney excised, the left renal pedicles were without any treatment. In IRI group, the pedicle of the left kidney was clamped for 30 min followed by various reperfusion periods (6, 12, 24 h). To study the effects of LSD1, TCP (MedChemExpress) was injected intraperitoneally at different doses (2.5, 5, 10 mg/kg) before IRI model establishment, once a day for 1 week. TCP powder was dissolved in dimethyl sulfoxide (DMSO). In the vehicle control group, equal amount of DMSO was injected intraperitoneally.

One million 786O cells with or without shTAZ were implanted subcutaneously into the healthy 8-week-old JAX NOD.CB17-PrkdcSCID-J mice; both male and female mice were used. Once tumor volume reached 120 mm³, mice were randomized into control or erastin treatment group. The vehicle (ORA-plus) or erastin (0.1 ml of 4 mg/ml erastin) was administrated by oral gavage twice daily for 20 days.

Wild-type SD rats were kept in the Animal Experiment Center of Southeast University. Experimental rats were divided into 4 groups (n = 6 per group). The method of establishing the I/R model was provided in supplementary material. Then, we covered the ligation with gel. In order to fully cover the infarcted area of the heart, we chose to inject about 300 uL of mimics + Gel at 23 mm below the left atrial appendage (about the ligation). In order to prevent excessive irradiation of tissue burns, we selected each irradiation for 2 min to control the body surface temperature for a total of 10 min of irradiation.

Adult male C57BL6 (C57) mice (8-12 weeks, 20-25 g) were purchased from the Animal Experiment Center of Wuhan University. All 64 mice were randomly divided into various groups by different treatments (n = 8). In sham group, after the right kidney excised, the left renal pedicles were without any treatment. In IRI group, the pedicle of the left kidney was clamped for 30 min followed by various reperfusion periods (6, 12, 24 h). To study the effects of LSD1, TCP (MedChemExpress) was injected intraperitoneally at different doses (2.5, 5, 10 mg/kg) before IRI model establishment, once a day for 1 week. TCP powder was dissolved in dimethyl sulfoxide (DMSO). In the vehicle control group, equal amount of DMSO was injected intraperitoneally.

One million 786O cells with or without shTAZ were implanted subcutaneously into the healthy 8-week-old JAX NOD.CB17-PrkdcSCID-J mice; both male and female mice were used. Once tumor volume reached 120 mm³, mice were randomized into control or erastin treatment group. The vehicle (ORA-plus) or erastin (0.1 ml of 4 mg/ml erastin) was administrated by oral gavage twice daily for 20 days.

2.5 x 10⁵ NCI-H1650 cells were inoculated 1:1 in Matrigel: PBS (100 mL) by subcutaneous injection into eight non-obese diabetic (NOD) severe combined immunodeficiency (SCID) gamma male mice. Tumors were allowed to engraft and grow for 30 days (tumor volume averaged ~200 mm³) and mice treated by intraperitoneal (i.p.) injection with 100 mg/kg cyst(e)inase or 100 mg/kg heat-inactivated cyst(e)inase (n = 4 ea.) on day 30, with a second dose given on day 33. Mice were necropsied 24 hr after the second dose.

Twenty athymic BALB/c nude mice (aged 4 weeks, weight 20-22 g, from Shanghai laboratory animal Center, Shanghai, China) were housed in a specific pathogen-free environment. A total of 1 x 10⁶ logarithmically growing C6 cells in 100 uL of PBS were subcutaneously injected into the right flank of each mouse. Therapeutic experiments were started when the tumor reached about 150 mm³ after about 7 days. The mice were allocated to receive intraperitoneal injections of vehicle (control group, n = 5/group), PAB at the dosage of 10 mg/kg body weight (n = 10/group) and 20 mg/kg body weight (n = 10/group) in the same volume 50 uL once a days for 8 times.

8-week C57BL/6J male mice purchased from Comparative Medicine Center of Yangzhou University were retained with unrestricted access to sterilized diet and water at standard bio-clean laboratory settings (Experimental Animal Center of College of Veterinary Medicine of Yangzhou University). Animals were randomly divided into four groups(n = 6-8 mice per group): control group or ISO group: injected with saline or ISO (5 mg/kg) subcutaneously for 14 days and, meanwhile, received vehicle saline via gavage for 14 days respectively; ATV (Pfizer,USA) group or ISO + ATV group: injected with saline or 5 mg/kg ISO (Sigma, USA) subcutaneously for 14 days and, meanwhile, received 20 mg/kg ATV via gavage for 14 days respectively.

Male Sprague Dawley ratsweighing 80-100 g were used to make the HF model induced by descending aortic banding (AB) procedure. Rats receiving a similar procedure except for the arterial ligation were defined as the sham-operated (SO) group. After the procedure, echocardiography was immediately applied to confirm the arterial banding. Rats receiving subcutaneous injections of low- or high-dose puerarin (100 mg/kg/day and 200 mg/kg/day, respectively) after the AB procedure were respectively defined as the Pue1 and Pue2 groups. An equal volume of normal saline was injected into the rats of the SO and AB groups.

We purchased forty-eight 3-week-old male C57BL/6N mice from Beijing HFK Bioscience Co. Ltd. and gave a twelve-hour light and dark cycle starting from 06:00 (am) to 18:00 (pm). Mice were randomly assigned into three groups after 2 weeks of adaptive feeding as follows. (1) The control group (n = 16): mice were provided with normal drinking water, a normal diet and intraperitoneal administration of solvent (5% DMSO + 40% Peg300 + 5% Tween 80 + 50% ddH2O) 3 mL/kg once a week aged 13 to 17 weeks. (2) The HFpEF group (n = 16): a double-hit model was designed, in which metabolic and mechanical stress worked together and resulted in HFpEF. Briefly, C57BL/6N mice had unrestricted access to a high-fat diet (HFD, D12492, Research Diet) starting from 5 weeks old. Meanwhile, a nitric oxide synthase inhibitor, N (gamma)-nitro-L-arginine methyl ester (L-NAME) (N5751, Sigma) was supplied in drinking water (0.5 g/L) for HFpEF groups, and the pH of the drinking water was adjusted to 7.4. The above placebo solvent was administrated in the same manner. (3) The HFpEF + Levo group (n = 16): according to the previous study, HFpEF mice received 3 mg/kg levosimendan (S2446, Selleck) (Dissolve 1 mg of levosimendan in 50 uL of DMSO, subsequently dilute to 1 mg/mL with the above solvent) intraperitoneally once a week from week 13 to 17.