Hsf1 and Hsf1+/+-/- mice were kindly given as a present by Dr. Ivor J. Benjamin (Froedtert & Medical College of Wisconsin, Milwaukee, WI, USA). Sex-matched Hsf1-/- mice and Hsf1 littermates were used at 16-20 weeks old. Each mouse was injected intraperitoneally with 2.5 umol PA (dissolved in 0.5 mL 10% BSA) or an equal volume of BSA twice daily for 7 days.

All mice used in this study were 6-12 weeks old on a C57/BL6/J background. WT or T-KO mice were fed with water supplemented with methionine (1 mgl-1, Sigma) or Se-Met (1 mgl-1, Sigma) and maintained on the diets for 4 weeks before experiments. Alternatively, WT mice were fed with selenium-adequate (0.15 mg/kg) and selenium-high (1 mg/kg) diets that were purchased from Envigo and mice were maintained on the diets for 4 weeks before experiments.

C57BL/6 WT mice were purchased from Beijing HFK BioScience Company (Beijing, China). Gpx4flox/flox mice were crossed with Vav-Cre mice and Mx-Cre mice to generate the Gpx4flox/flox Vav-Cre mice and Gpx4flox/flox Mx-Cre mice, respectively. For Gpx4 deletion, Gpx4flox/flox Mx-Cre mice were intraperitoneally injected with 20 mg/kg pIpC (Sigma) every other day for two weeks. CD45.1/45.2 mice and CD45.1 mice on a C57BL/6 background were used as competitor and recipient mice, respectively, in the competitive transplantation assay. Mice were fed natural ingredient diets containing >= 120 IU/kg vitamin E. A fixed formulation diet with or without 75 IU/kg vitamin E (Beijing HFK BioScience Company, Beijing, China) was fed to the mice involved in the vitamin E depletion experiments. For 5-FU treatment, mice were intraperitoneally injected with 150 mg/kg 5-FU (Sigma).

Male healthy Wistar rats (six-week-old, provided by Experimental Animal Centre of Harbin Medical University, China) were used in this study. All rats (3-4 rats per cage) access to standard diet anddeionized waterad libitum and were placed in standard laboratory conditions. Seventy-two rats (weighing 200-220 g) were randomly divided into 4 groups (n = 18): AlNPs group was exposed to 50 mg/kg AlNPs (< 50nm, Sigma-Aldrich, USA) by gavage once a day for 90 days. CRS + AlNPs group was received CRS for 21 days and was exposed to 50 mg/kg AlNPs daily by gavage for 90 days. CRS + H2O group was subjected to CRS for 21 days and was given the same volume of deionized water daily by gavage for 90 days. The control (CON) group was given the same volume of deionized water daily and not affected by restraint stress for 90 days.

Male healthy Wistar rats (six-week-old, provided by Experimental Animal Centre of Harbin Medical University, China) were used in this study. All rats (3-4 rats per cage) access to standard diet anddeionized waterad libitum and were placed in standard laboratory conditions. Seventy-two rats (weighing 200-220 g) were randomly divided into 4 groups (n = 18): AlNPs group was exposed to 50 mg/kg AlNPs (< 50nm, Sigma-Aldrich, USA) by gavage once a day for 90 days. CRS + AlNPs group was received CRS for 21 days and was exposed to 50 mg/kg AlNPs daily by gavage for 90 days. CRS + H2O group was subjected to CRS for 21 days and was given the same volume of deionized water daily by gavage for 90 days. The control (CON) group was given the same volume of deionized water daily and not affected by restraint stress for 90 days.

Male healthy Wistar rats (six-week-old, provided by Experimental Animal Centre of Harbin Medical University, China) were used in this study. All rats (3-4 rats per cage) access to standard diet anddeionized waterad libitum and were placed in standard laboratory conditions. Seventy-two rats (weighing 200-220 g) were randomly divided into 4 groups (n = 18): AlNPs group was exposed to 50 mg/kg AlNPs (< 50nm, Sigma-Aldrich, USA) by gavage once a day for 90 days. CRS + AlNPs group was received CRS for 21 days and was exposed to 50 mg/kg AlNPs daily by gavage for 90 days. CRS + H2O group was subjected to CRS for 21 days and was given the same volume of deionized water daily by gavage for 90 days. The control (CON) group was given the same volume of deionized water daily and not affected by restraint stress for 90 days.

Male healthy Wistar rats (six-week-old, provided by Experimental Animal Centre of Harbin Medical University, China) were used in this study. All rats (3-4 rats per cage) access to standard diet anddeionized waterad libitum and were placed in standard laboratory conditions. Seventy-two rats (weighing 200-220 g) were randomly divided into 4 groups (n = 18): AlNPs group was exposed to 50 mg/kg AlNPs (< 50nm, Sigma-Aldrich, USA) by gavage once a day for 90 days. CRS + AlNPs group was received CRS for 21 days and was exposed to 50 mg/kg AlNPs daily by gavage for 90 days. CRS + H2O group was subjected to CRS for 21 days and was given the same volume of deionized water daily by gavage for 90 days. The control (CON) group was given the same volume of deionized water daily and not affected by restraint stress for 90 days.

Sprague-Dawley rats (4 weeks) were obtained from the Animal Center of the Guangzhou University of Chinese Medicine. Procurement, maintenance, and treatment of the animals were performed under the supervision of the Institutional Animal Ethics Committee of the Guangzhou University of Chinese Medicine. The Gaussian 16 C.01 program was purchased from Guangzhou Molcalx Ltd. (Guangzhou, China).

Sprague-Dawley rats (4 weeks) were obtained from the Animal Center of the Guangzhou University of Chinese Medicine. Procurement, maintenance, and treatment of the animals were performed under the supervision of the Institutional Animal Ethics Committee of the Guangzhou University of Chinese Medicine. The Gaussian 16 C.01 program was purchased from Guangzhou Molcalx Ltd. (Guangzhou, China).

The C. elegans strain N2 was grown on NGM plates and on OP50 bacteria as a food source. For the lactate measurement, the nematodes were synchronized by bleaching with NaOH/NaClO and grown on NGM plates containing OP50 and either DMSO, 100 uM CyPPA or 250 uM CyPPA for 4 days. To determine lactate levels, the lactate assay kit was used (Sigma-Aldrich, MAK064). Briefly, worms were collected and washed to remove OP50. The worm pellet was snap-frozen, resuspended in lactate assay buffer and sonicated to break the cuticle. After centrifugation at 13000 rpm for 10 min, the supernatant was applied to a 10 kDa MWCO filter and centrifuged for 60 min at 13000 rpm to remove insoluble material.

Sprague-Dawley rats were purchased from the Animal Centre of Guangzhou University of Chinese Medicine. male Sprague-Dawley rats were collected, and the adherent soft tissues were removed. Both ends of the bones were cut away from the diaphysis with bone scissors. The bone marrow plugs were hydrostatically expelled from the bones by insertion of needles fastened to 10-mL syringes filled with complete medium; the needles were inserted into the distal ends of femora and proximal ends of the tibiae, and the marrow plugs expelled from the opposite ends.

Sprague-Dawley rats were purchased from the Animal Centre of Guangzhou University of Chinese Medicine. male Sprague-Dawley rats were collected, and the adherent soft tissues were removed. Both ends of the bones were cut away from the diaphysis with bone scissors. The bone marrow plugs were hydrostatically expelled from the bones by insertion of needles fastened to 10-mL syringes filled with complete medium; the needles were inserted into the distal ends of femora and proximal ends of the tibiae, and the marrow plugs expelled from the opposite ends.

A synchronous population was obtained by transferring egg-laying adults to fresh plates at 16 for 2-3 hr. The adults were removed and the plates with eggs then transferred to 25 to ensure sterility. After 48 hr at 25, when worms were at the late L4/young adult stage, 25-35 nematodes were transferred to fresh plates containing either vehicle control, 250 uM SIH, or 200 uM Lip-1. All plates were coded to allowing blinding of the experimenter to the treatment regime during scoring.

Male 20 weeks old CD-1 mice were obtained from Charles River Laboratories. The mice were randomly assigned (using random number generator in Excel) to receive 5 mg/kg Fer-1 (Abcam, cat #ab146169) or 1.5% DMSO (vehicle) 60 minutes before injection of folic acid (Sigma-Aldrich) of 250 mg/kg in 0.3 mol/L sodium bicarbonate intraperitoneally. Mice were euthanized 48 h later.

Male 20 weeks old CD-1 mice were obtained from Charles River Laboratories. The mice were randomly assigned (using random number generator in Excel) to receive 5 mg/kg Fer-1 (Abcam, cat #ab146169) or 1.5% DMSO (vehicle) 60 minutes before injection of folic acid (Sigma-Aldrich) of 250 mg/kg in 0.3 mol/L sodium bicarbonate intraperitoneally. Mice were euthanized 48 h later.

All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pennsylvania. Mice were maintained on a mixed C57BL/6 background on a standard light-dark cycle. Mice carrying Mll4SET floxed alleles, Mll3SET floxed alleles, or a combination of both of these were crossed with Krt14-Cre transgenic mice.

PDX models of triple-negative breast cancer were obtained from the Dana-Farber Cancer Institute and propagated in NSG mice. Tumors were harvested and digested using collagenase at 37. Once digested, the cells were filtered using a cell strainer (40 um), washed twice with PBS, and plated in DMEM/F12 (containing 10% FBS).

PDX models of triple-negative breast cancer were obtained from the Dana-Farber Cancer Institute and propagated in NSG mice. Tumors were harvested and digested using collagenase at 37. Once digested, the cells were filtered using a cell strainer (40 um), washed twice with PBS, and plated in DMEM/F12 (containing 10% FBS).

All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pennsylvania. Mice were maintained on a mixed C57BL/6 background on a standard light-dark cycle. Mice carrying Mll4SET floxed alleles, Mll3SET floxed alleles, or a combination of both of these were crossed with Krt14-Cre transgenic mice.

5.0 x 10⁶ cells were mixed with Matrigel (BD Biosciences) at 1:1 ratio (v/v) and injected subcutaneously into seven-week old nude mice (NU/NU; Charles River). Mice were fed with regular chow. After nine weeks, the mice were killed and the tumors were weighed and recorded.

5.0 x 10⁶ cells were mixed with Matrigel (BD Biosciences) at 1:1 ratio (v/v) and injected subcutaneously into seven-week old nude mice (NU/NU; Charles River). Mice were fed with regular chow. After nine weeks, the mice were killed and the tumors were weighed and recorded.

All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pennsylvania. Mice were maintained on a mixed C57BL/6 background on a standard light-dark cycle. Mice carrying Mll4SET floxed alleles, Mll3SET floxed alleles, or a combination of both of these were crossed with Krt14-Cre transgenic mice.