EMBO/soda-nel huggingface dataset:
Calcineurin promotes APC/C activation at meiotic exit by acting on both XErp1 and Cdc20 (2018)
Figure 3

Figure 3 Thr68 of Cdc20 is a target site of calcineurin.

A) CSF extract was supplemented with mRNA encoding Flag-Cdc20 that was either wild-type or mutated to serine at Thr64, Thr68 and Thr79 (3TS). After expression, calcium was added, samples were taken at the indicated time points and resolved by either conventional or Phos-tagTM SDS-PAGE. Samples were immunoblotted for the Flag-tag, Cdc27 and cyclin B2.

B) CSF extract was supplemented with Myc-XErp1 wt IVT and treated with DMSO or CsA. Calcium was added, samples were taken at the indicated time points, resolved by either conventional or Phos-tagTM SDS-PAGE and immunoblotted for Cdc20, Cdc27 and cyclin B2. A control sample was treated with λ-phosphatase. A pseudocoloured representation of the Cdc20 and the Cdc27 immunoblots is shown in Fig. EV4A. The cyclin B2 membrane was stripped and reprobed for α-tubulin.

C) CSF extract was supplemented with Flag-Cdc20 wt IVT coupled to α-Flag beads. Calcium was added and Flag-Cdc20 was isolated by α-Flag immunoprecipitation at the indicated time points. Input and pellet (IP: Flag) samples were immunoblotted for the Flag-tag, pThr68 Cdc20 and p150(Glued). The pThr68 Cdc20 membrane was stripped and reprobed for cyclin B2. Asterisk in the cyclin B2 immunoblot indicates the IgG heavy chain. Asterisks in other immunoblots indicate unspecific bands.

D) CSF extract was supplemented with Flag-Cdc20 wt IVT. Flag-Cdc20 was re-isolated by α-Flag immunoprecipitation, supplemented with calcium and treated with recombinant hs_CaN and calmodulin as indicated. Samples were taken at the indicated time points and immunoblotted for the Flag-tag and pThr68 Cdc20. The pThr68 Cdc20 membrane was stripped and reprobed for the catalytic calcineurin subunit CnA. Asterisk indicates the IgG heavy chain.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 1

Figure 1: VEGF-A is strongly expressed in the RPE, the lens and the ciliary body in the eye, and its expression is maintained with progressive age.

A.-E. Staining for β-galactosidase in 3-months old VEGF-Ahyper (lacZ/wt) mouse eyes.

A. Low magnification image shows β-gal+ cells (in blue) in the RPE (white arrow), retina (blue arrow), ciliary body (black arrow), and lens (orange arrow). Images of Figures 1B to 1E are higher magnification images of Figure 1A. Scale bar 200 μm.

B. Strong expression of VEGF with β-gal+ cells are observed in the RPE (short white arrow) and the inner nuclear layer (INL) (long white arrow), while less staining is observed in the ganglion cell layer (black arrow) of the retina. Scale bar 100 μm.

C. Ciliary body epithelial cells are β-gal+, similarly as RPE cells (white arrow). Scale bar 100 μm.

D. VEGF-A is strongly expressed in secondary nuclear fiber cells in the adult lens (white arrow). Scale bar 50 μm.

E. No β-gal+ cells are observed in the corneas of adult VEGF-Ahyper (lacZ/wt) mouse eyes. White arrow shows the stroma of the cornea and the black arrow shows the corneal epithelium. Scale bar 100 μm.

F. No staining for β-galactosidase is observed in 3-months old control wild-type littermate eyes. Scale bar 200 μm.

G. RPE cells maintain strong expression of VEGF-A (β-gal+ cells [white arrow]) in the retina with progressive age. Immunofluorescence for β-gal in the retina of a 24-months old VEGF-Ahyper mouse is shown. Ocular macrophages (F4/80+ cells [yellow arrow]) show no strong β-galactosidase expression, while some immunolabeling for β-galactosidase is detected in retinal cells of the inner nuclear layer (orange arrow) of the retina. RPE: retinal pigment epithelium; ONL: outer nuclear layer; INL: inner nuclear layer. Scale bar 50 μm.

H. VEGF-A continues to be highly expressed in secondary nuclear fiber cells in the aged lens (shown is a lens of a 24-months old VEGF-Ahyper mouse; nuclear β-galactosidase staining (arrow) reflects cellular VEGF-A expression). Cor: lens cortex; Cap: lens capsule. DAPI labels nuclei (blue). Scale bar 25 μm.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 2

Figure 2: Increased VEGF-A expression and 4-HNE levels in aged VEGF-Ahyper mice.

A. Left graph: VEGF-A protein levels in lenses of VEGF-Ahyper mice (KI) are increased ~3-fold throughout life (measured at 2- and at 8.5-months of age; n=7 mice/group, 2 independent experiments). Graph shows mean ± SD. Right graph: VEGF-A mRNA levels are similarly increased in lenses of adult VEGF-Ahyper mice (9-months old; n=7 mice/group, 3 independent experiments). Graph shows mean ± SEM. **P-value: 0.0095.

B. VEGF-A isoforms (mainly VEGF-A120 and VEGF-A164) and the VEGF-A receptor Flk1 are expressed in the adult lens. Flt1 expression was not detected in the lens. N=3 mice/group.

C. Increased serum levels of the lipid peroxidation marker 4-HNE (a marker of increased oxidative damage) in aged, but not in young VEGF-Ahyper mice. *P-value: 0.0325. N=7 mice/group (2 independent experiments). Graph shows mean ± SEM.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 3

Figure 3: Higher VEGF-A levels in the lens cause ERK-pathway hyperactivation and age-dependent progressive nuclear cataracts.

A. Increased ERK1/2 phosphorylation is observed in lenses from VEGF-Ahyper mice (KI) with increased VEGF-A levels already prior to cataract formation (11-months old mice with no cataracts (0 cataract)), which is maintained in lenses with mature cataracts (lens of a 26-months old VEGF-Ahyper mouse with +3 cataract). Quantification of ERK1/2 phosphorylation in 11-months old lenses (n=3/group). Graph shows mean ± SEM. ***P-value: 0.0005.

B.-C. Cataracts with opacification of the lens (B., black arrow) and extruded nuclei (C., white arrow) are observed with progressive age in VEGF-Ahyper mice (here a representative 30-months old lens with a cataract is shown).

D.-E. Histology of lens from b.-c. shows extrusion of sclerotic nucleus (white arrows in C. and D.) and massive degenerative changes of the lens cortex with vacuolization (black arrows in D. and E.). Scale bar 500 μm in (D.) and 100 μm in (E.).

F.-G. Representative histological images of 21-months old eyes from WT and VEGF-Ahyper mice. The sclerotic nucleus (red arrow in G.) was extruded and the lens cortex and capsule are separated from the nucleus (white arrow in G.). The retina in aged VEGF-Ahyper mice is atrophic and shows degenerative changes (yellow arrow in G.), while the retina (*) and lens (**) appears normal in age-matched WT littermate mice. Scale bars 200 μm. DAPI (blue, nuclei); phalloidin (green).
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 4

Figure 4: Increased lenticular VEGF-A increases markers of oxidative stress in the lens and targeting NLRP3 delays cataract formation.

A. Cataracts progress with age in VEGF-Ahyper mice. Cataracts were graded from +1 (mild), +2 (moderate), to +3 (mature cataract with fully opacified lens). While only a small subset of >18 months-old WT mice have mature (+3) cataracts, the majority of VEGF-Ahyper mice of this age group has mature cataracts. Percentile (%) of mice with graded cataracts is indicated in each age group. 0 = no cataract. Absolute mouse numbers of each group are shown in Appendix Figure S1.B. Targeting NLRP3 inhibits normal age-dependent cataract formation, and delays (but does not prevent) VEGF-A-induced cataract formation in VEGF-Ahyper mice. Percentile (%) of mice with graded cataracts is indicated in each age group. 0 = no cataract. Absolute mouse numbers of each group are shown in Appendix Figure S1.

C. Expression of IL-1β in lenses of 21-months old VEGF-Ahyper mice with +3 cataracts is increased compared to lenses from age-matched WT mice with +3 cataracts, but not in lenses from 11-months old mouse groups with no cataracts. *P-value: 0.0259. N=7/group (3 independent experiments).

D. VEGF-Ahyper mice have decreased levels of the major antioxidant in the lens, reduced glutathione (μmol/gm lens weight), prior to cataract formation. Lenses with 0 to +1 cataracts from 11-months old mice were used for measurements. ** P-value: 0.0097. N=7/group (2 independent experiments).

E-F. Overexpression of NADPH oxidase gp91phox [*p-value: 0.0435] (E.) and of the antioxidant enzymes SOD1 [*p-value: 0.0330], catalase (CAT) [*p-value: 0.0448] and GPx-1 [*p-value: 0.0443] (F.) in 21-months old lenses of VEGF-Ahyper mice with cataracts (+3), but not in 11-months old lenses with no cataracts. N=7/group (3 independent experiments).

Graphs show mean ± SEM.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 5

Figure 5: Choroidal neovessels originate from the choroidal vasculature in VEGF-Ahyper mice.

A-B. Choroidal flat mount staining of a white VEGF-Ahyper mouse shows choroidal vessels (green [fluorescein-conjugated isolectin B4]) from which neovessels (strong red [CD31]) originate and that are covered by a RPE cell layer (white [phalloidin]), which separates the CNV lesion from the photoreceptors. White arrows demarcate CNV lesion. Yellow arrows show round autofluorescent deposits. Scale bars 100 μm.

C. Section through a CNV lesion in a VEGF-Ahyper mouse. Bruch's membrane location is indictaed by yellow arrows. Choroidal neovessels (white arrows) are covered by a layer of RPE cells (black arrows). Chor: choroid; ONL: outer nuclear layer; PR: photoreceptors. Scale bar 20 μm. A.-C. 21-months old VEGF-Ahyper mice.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 6

Figure 6: Increased VEGF-A in the RPE causes neovascular AMD-like pathologies by inducing RPE barrier breakdown through activation of Flk1-signaling in the RPE.

RPE-specific overexpression of VEGF-A in VMD2Cre+/WTROSA-STOPfl/fl-VEGF-A164 mice (VEGF-ARPE mice) results in CNV at sites of VEGF-A overexpression (Cre+ patches (white nuclei); red arrow), while Cre- areas with normal VEGF-A expression show no CNV (white arrow). Neovessels are CD31+ (red). VEGF-A-induced CNV lesions and RPE barrier breakdown in VMD2Cre+/WT ROSA-STOPfl/fl-VEGF-A164 mice resemble those seen in VEGF-Ahyper mice (B.-C., arrows; phalloidin staining in green). Scale bars 100 μm.

D. Genetic inactivation of Flk1 specifically in the RPE in VEGF-Ahyper mice (VMD2Cre+/WTFlk1fl/fl VEGF-Ahyper mice) prevents RPE barrier breakdown and CNV lesion formation. Choroidal flat mounts of 5 months old VEGF-Ahyper mice and VMD2Cre+/WTFlk1fl/fl VEGF-Ahyper mice are shown. β-gal staining shows uniform VEGF-A expression in these mice (green). Cre staining (red) shows co-localization of β-gal and Cre staining in the nuclei of RPE cells in VMD2Cre+/WTFlk1fl/fl VEGF-Ahyper mice (red areas of staining occur in CNV lesions of VEGF-Ahyper mice due to labeling of anti-mouse IgG and not Cre, as the anti-mouse IgG secondary antibodies were used to detect the mouse anti-Cre antibody). Phalloidin staining (white) reveals extensive RPE barrier breakdown and CNV lesions (arrow) in VEGF-Ahyper mice, which is not observed in VMD2Cre+/WTFlk1fl/fl VEGF-Ahyper mice. Scale bar 100 μm.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 7

Figure 7: Increased NLRP3, complement C1q and C5b-9 in CNV lesions of VEGF-Ahyper mice.

A. NLRP3 is expressed in RPE cells at sites of CNV lesions (arrow; white).

B. Lack of NLRP3 staining in VEGF-Ahyper/NLRP3-/- mice (control). Scale bars 100 μm.

C. Expression of NLRP3 [*p-value: 0.0478] and IL-1β [*p-value: 0.0398], but not of IL-18 [p-value: 0.3529] is increased in RPE/choroid lysates from VEGF-Ahyper mice when compared to matched control littermate mice. N=7 mice/group (3 independent experiments). Graphs show mean ± SEM.

D.-G. Complement pathway activation (immunolabeling for the product of complement pathway activation C5b-9, an inducer of the NLRP3 inflammasome) is observed in CNV lesions of VEGF-Ahyper mice (white arrow in E.) but not in age-matched control mice (D.) Co-localization of C5b-9 in the same CNV lesion is observed with complement C1qA (the initiator of the classical complement pathway and an inducer of the NLRP3 inflammasome) (F.), and with TLR2 (G.) (white arrows). Autofluorescence in red in D. and E. CD31+ vessels in red in I. and J. Yellow arrows show site of choroid. Scale bars 50 μm.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 8

Figure 8: VEGF-A-induced AMD-like pathologies occur in an NLRP3 inflammasome- and IL-1β-mediated process, which is regulated by TLR2-signaling and autophagy pathways.

A. CNV lesion numbers per eye in 6-week old VEGF-Ahyper mice compared with VEGF-Ahyper mice lacking CASP1/CASP11, NLRP3, IL1R1, IL-18, TLR2, or those heterozygous for beclin-1 or homozygous for hypomorphic ATG16L1. CASP1/CASP11, NLRP3 or IL-1R1 deficiency results in significantly fewer CNV lesions. Each value represents the total CNV lesion number/eye for an individual mouse. Mean ± SEM is shown. P-values are based on comparison with the VEGF-Ahyper mouse group. Absolute numbers of mice per group are indicated in ( ). P-values were calculated with a two-tailed unpaired Student’s t-test. Separately, p-values are shown between VEGF-Ahyper/CASP1-/-/CASP11-/- and VEGF-Ahyper/NLRP3-/- or VEGF-Ahyper/IL1R1-/- mouse groups. WT mice developed no CNV lesions.

B. Average CNV lesion area (in μm2/lesion) in 6-week old VEGF-Ahyper mice compared with VEGF-Ahyper mice lacking CASP1/CASP11, NLRP3, IL1R1, IL-18, TLR2, or those heterozygous for beclin-1 or homozygous for hypomorphic ATG16L1. Lack of TLR2 or CASP1/CASP11 significantly reduces CNV lesion size, while reduced autophagy moderately increases CNV size. Each value represents the average CNV lesion size/eye for an individual mouse. Mean ± SEM is shown. P-values are based on comparison with the VEGF-Ahyper mouse group. Absolute numbers of mice per group are indicated in ( ). P-values were calculated with a two-tailed unpaired Student’s t-test. Separately, p-values are shown between VEGF-Ahyper/CASP1-/-/CASP11-/- and VEGF-Ahyper/NLRP3-/- or VEGF-Ahyper/IL1R1-/- mouse groups.

C.-E. Representative choroidal flat mount images show multifocal CNV lesions in VEGF-Ahyper mice and a significant reduction in CNV numbers in those VEGF-Ahyper mice that lack either CASP1/CASP11 or IL1R1 (and therefore IL-1β-signaling). Staining for phalloidin in green and for CD31 in red. Scale bars 200 μm.

F. Laser-induced CNV experiments demonstrate that treatment with a caspase-1 inhibitor (compared to DMSO treated controls) or a neutralizing TLR2 antibody (compared to isotype matched IgG treated controls) inhibits CNV lesion growth. P-values are shown. N=15 mice/group. Representative CNV lesions in choroidal flat mounts are shown. Scale bars 100 μm.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 9

Figure 9: VEGF-A-induced non-exudative AMD-like pathologies can be prevented by targeting NLRP3, IL1R1 or caspase-1/caspase-11.

A. No significant abnormalities of the choroid, RPE or retina are seen in control wild-type littermate mice. Representative image of a 28-months old control eye is shown. White arrow indicates normal Bruch's membrane. Chor: choroid; RPE; PR: photoreceptor; ONL: outer nuclear layer.

B. Aged VEGF-Ahyper mice develop a progressive age-dependent degeneration of the RPE (red arrow) and the photoreceptors (yellow arrow) with accumulation of thick sub-RPE basal laminar deposits (black arrow). These non-exudative AMD-like pathologies occur also at sites devoid of CNV lesions.

C.-E. Genetic inactivation of CASP1/CASP11, NLRP3, or IL1R1 strongly inhibits the manifestation of RPE and photoreceptor degeneration in aged VEGF-Ahyper mice.

F.-G. VEGF-Ahyper/IL18-/- mice show significant RPE and photoreceptor degeneration, with shortened and irregular photoreceptor outer segments (white arrows). Thick sub-RPE deposits that protrude into the RPE plane are observed as well (black arrows).

H. VEGF-Ahyper/Becn1+/- mice show significant RPE and photoreceptor degeneration.

I. In contrast, age-matched VEGF-Ahypo mice that are hypomorphic for VEGF-A and express β-galactosidase from the endogenous VEGF-A locus (as do VEGF-Ahyper mice) do not show these eye pathologies.

B-I. 21-months old mice. Scale bars 20 μm.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 10

Figure 10: Targeting caspase-1/caspase-11 or IL1R1 rescues loss of visual function in aged VEGF-Ahyper mice.

A. Graphs show means of a- and b-waves of dark-adapted and light-adapted ERG responses (ERG intensities in μV) for both the left and the right eyes from 21-months old VEGF-Ahyper mice, VEGF-Ahyper/CASP1-/-/CASP11-/- mice, CASP1-/-/CASP11-/- mice, VEGF-Ahyper/IL1R1-/- mice, IL1R1-/- mice, VEGF-Ahyper/IL18-/- mice, and IL18-/- mice. None of these mice had cataracts. Graphs show mean ± SEM.

P-values relative to the VEGF-Ahyper mouse group are indicated.

B. Representative ERG tracings are shown for VEGF-Ahyper mice and for VEGF-Ahyper/CASP1-/-/CASP11-/- mice. X-axes show milliseconds (ms). Y-axes show ERG intensities (in μV).

C. For each experimental mouse group means for a- and b-waves of ERGs were calculated and p-values are shown for left and right eyes using a two-tailed unpaired Student's t-test.

P-values of the VEGF-Ahyper mouse group are compared to those VEGF-Ahyper mice lacking either CASP1/CASP11, IL1R1 or IL18, as well as to the KO mice (CASP1-/-/CASP11-/- mice, IL1R1-/- mice, or IL18-/- mice) without the VEGF-Ahyper allele. In addition, p-values are shown comparing values between to KO mouse groups and those that have the VEGF-Ahyper allele. Mouse numbers for each experimental mouse group is indicated as well.
Increased VEGF-A promotes multiple distinct aging diseases of the eye through shared pathomechanisms (2016)
Figure 11

Figure 11: Lack of caspase-1/caspase-11 inhibits CNV lesion formation, retinal degeneration, fundus abnormalities and fundus autofluorescence in aged VEGF-Ahyper mice.

A. OCTs in aged VEGF-Ahyper mice show retinal degeneration with a thinning of the retina and CNV lesions extending from the choroid into the subretinal space (arrows). OCTs appear normal in age-matched VEGF-Ahyper/CASP1-/-/CASP11-/- mice and CASP1-/-/CASP11-/- mice.

B. Fundus autofluorescence imaging shows areas of autofluorescence in the fundus of aged VEGF-Ahyper mice, but not in age-matched VEGF-Ahyper/CASP1-/-/CASP11-/- mice and CASP1-/-/CASP11-/- mice.

C. Abnormal pigment loss and degenerative changes are observed in the fundus of aged VEGF-Ahyper mice, while the fundus of age-matched VEGF-Ahyper/CASP1-/-/CASP11-/- mice and CASP1-/-/CASP11-/- mice does not show these abnormalities.

Representative images of 21-months old mice are shown.
The mitochondrial Ca2+ uptake regulator, MICU1 is involved in cold stress-induced ferroptosis (2021)
Figure 2

Figure 2. DIME interaction and dimerization domains of MICU1 are necessary for cold stress-induced ferroptosis

(A) Schematic model describing the domains of MICU1 and the brief structure of the MCU complex green points describing Ca2+ and red points describing mutants.

(B) Immunoblots of MICU1 and its mutants rescued by the lentivirus. Arrows indicate MICU1 or the dimer. * indicates the potential DTT-resistant dimer.

(C) LDH assay was performed after 16 h of cold stress on ice. Data are presented as mean ± SEM; n = 3, biological replicates. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, n.s.: not significant, by one-way ANOVA followed by Tukey's multiple comparison test

(D, E) Lipid peroxidation was measured by C11-BODIPY 581/591 after cold stress for 5 h. Representative plots are shown in (D). Quantification data (E), whose raw value were normalized by the C11-BODIPY 581/591 of the WT A549 cells after cold stress application, are presented as mean ± SEM; n = 4, biological replicates. ****p < 0.0001, ***p < 0.001, **p < 0.01, n.s.: not significant, by one-way ANOVA followed by Tukey's multiple comparison test.
The mitochondrial Ca2+ uptake regulator, MICU1 is involved in cold stress-induced ferroptosis (2021)
Figure 3

Figure 3. Cold stress-induced mitochondrial membrane potential hyperpolarization contributes to lipid ROS accumulation and ferroptosis

(A) Schematic model of the electron transport chain (ETC) and targets of the inhibitors used.

(B-F) MMP was monitored by TMRE fluorescence at every 1 h. FCCP (200 µM) and antimycin A (50 µM) were used as pretreatments for 30 min. Data were normalized by basal state of WT A549 cells (B-D) or WT A549 cells with DMSO (E-F) before cold stress. Data at 1 h (C), 5 h (D, F) are presented as mean ± SEM; n = 4 (B-D) and n = 5 (E-F), biological replicates, ****p < 0.0001, **p < 0.01, *p < 0.05 by paired T-test (C), or one-way ANOVA followed by Dunnett's multiple comparison test (D, F).

(G-J) (G, J) LDH release was measured after cold stress for 24 h. FCCP (200 µM), antimycin A (50 µM), mitoQ and decylQ (500 nM) were used as pretreatments for 30 min. Data are presented as mean ± SEM; n = 4 (G) and n = 3 (J), biological replicates, ****p < 0.0001, ***p < 0.001, **p < 0.01, by one-way ANOVA followed by Dunnett's or Bonferroni's multiple comparison test. (H, I) Lipid peroxidation was measured by C11-BODIPY 581/591 after cold stress for 5 h. Representative plots are shown in (H). Quantification data (I), whose raw value were normalized by the C11-BODIPY 581/591 without cold stress of non-treatment, are presented as mean ± SEM; n = 5, biological replicates. **p < 0.01, *p < 0.05, by one-way ANOVA followed by Dunnett's multiple comparison test.
The mitochondrial Ca2+ uptake regulator, MICU1 is involved in cold stress-induced ferroptosis (2021)
Figure 4

Figure 4. Cytosolic Ca2+ is involved in cold stress-induced ferroptosis

(A) LDH release was measured after cold stress for 24 h. BAPTA-AM (10 µM) was used as pretreatments for 30 min. Data are presented as mean ± SEM; n = 3, biological replicates, ****p < 0.0001, by unpaired T-test.

(B-F) (B-C, E-F) Lipid peroxidation was measured by C11-BODIPY 581/591 after cold stress for 5 h. Representative plots are shown in (B, E). Quantification data (C, F), whose raw value were normalized by the C11-BODIPY 581/591 without cold stress of non-treatment, are presented as mean ± SEM; n = 4 (C), 3 (F), biological replicates. ***p < 0.001, **p < 0.01 by unpaired T-test. (D) LDH release was measured after cold stress for 16 h. Data are presented as mean ± SEM; n = 5, biological replicates, *p < 0.05, by unpaired T-test.

(G) Brief strategy for finding regulators for cold stress-induced ferroptosis and summary of molecular mechanism of lipid peroxidation mediated by MICU1.

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