EMBO/soda-nel huggingface dataset:
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 4

Figure 4. MAC assembly via surface-bound C5 convertases leads to inner membrane damage.

(A) Outer membrane damage (mCherry intensity) and inner membrane damage (% Sytox positive) of convertase-labeled perimCherry/cytoGFP E. coli cells incubated with a concentration range of C5 and fixed concentrations of C6-C9.

(B) Inner membrane damage of perimCherry/cytoGFP E. coli exposed to a concentration range of ΔC5 serum and, after washing, to C5-C9. As controls, bacteria were incubated with heat inactivated ΔC5 serum or 5 µM compstatin was added to the ΔC5 serum to block C3b deposition.

(C) Inner membrane damage of three different convertase-labeled bacteria exposed to buffer (Conv) or C5-C9 (Conv-MAC).

(D) Confocal microscopy images of convertase-labeled perimCherry/cytoGFP E. coli exposed to buffer (Conv) or C5-C9 (Conv-MAC). Unlabeled bacteria exposed to 1% serum served as control. Green = GFP, Red = To-pro-3 DNA dye. Scale bars = 3 µm.

Data information: (A-C) Data represent mean ± SD of 3 independent experiments. (C) Statistical analysis was done using a ratio paired two-tailed t-test and displayed only when significant as * p≤0.05 or ** p≤0.01.
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 5

Figure 5. Local assembly of C5b6 by surface-bound C5 convertases is required for killing.

(A) Schematic overview of MAC assembly on convertase labeled bacteria by C5b6 that is locally generated by incubation with C5 and C6 (top) or by preassembled C5b6 (bottom).

(B) Bacterial viability of convertase-labeled E. coli MG1655 exposed to Buffer (Conv), preassembled C5b6 (Conv + C5b6MAC) or a mixture of C5 and C6 (Conv-MAC), in the presence of C7, C8 and C9. Dotted line represents the detection limit of the assay.

C and D (C) Inner membrane damage (% Sytox positive) and (D) outer membrane damage (mCherry) of convertase-labeled perimCherry/cytoGFP E. coli exposed to a concentration range of preassembled C5b6 or a mixture of C5 and C6, in the presence of 100 nM C7. After washing, bacteria were exposed to 20 nM C8 and 100 nM C9.

Data information: (B-D) Data represent mean ± SD of 3 independent experiments. (B) Statistical analysis was done using a ratio paired two-tailed t-test and displayed only when significant as ** p≤0.01.
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 6

Figure 6. C5b6 rapidly loses the capacity to form bactericidal pores.

A, B Step-wise assembly of MAC on convertase-labeled bacteria. Convertase-labeled bacteria were incubated with C5/C6 or C5/C6/C7 for 15 minutes, and subsequently washed (@) or treated with 10 μg/ml Eculizumab (Ecu). Then, the remaining MAC components (C7-9 for C5/C6 or C8-9 for C5/C6/C7 respectively) were added to the incubation mixture. In the control conditions (Conv-MAC), the remaining MAC components were added to the incubation mixture without washing or adding an inhibitor. (A) Outer membrane (mCherry) and (B) inner membrane damage (% Sytox positive) were determined.

Data information: (A-B) Data represent mean ± SD of 3 independent experiments. Statistical analysis was done using a ratio paired two-tailed t-test in which the test conditions were compared to Conv-MAC and displayed only when significant as * p≤0.05.
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 7

Figure 7. Inner membrane damage is driven by MAC assembly at the outer membrane.

(A) Outer and inner membrane damage of convertase-labeled bacteria exposed to different combinations of MAC components. '@' indicates a washing step.

(B) Outer and inner membrane damage of convertase-labeled bacteria exposed to C5-C8 and after washing, to a concentration range of C9.

Data information: Data represent mean ± SD of 3 independent experiments. (A) Statistical analysis was done using a one-way ANOVA and displayed only when significant as ** p≤0.01 or **** p≤0.0001.
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 8

Figure 8. Local formation of C5b6 is required for efficient insertion of MAC pores into the outer membrane.

Surface-bound MAC pores were quantified by flow cytometry or confocal microscopy.

(A) Convertase-labeled bacteria were exposed to a concentration range of either preassembled C5b6 (C5b6MAC) or a mixture of C5 and C6 (Conv-MAC), in the presence of 100 nM C7. After washing, 20 nM C8 and 100 nM C9-Cy3 was added. Controls at 0 nM C5b6 or C5-C6 confirm that the detected C9-Cy3 deposition is specifically related to MAC formation (solid lines). Proper insertion of pores was assessed by a previously described shaving method with trypsin (Moskovich & Fishelson, 2007). Bacteria were first incubated with MAC components for 30 minutes and subsequently treated with 20 µg/ml trypsin for 15 min at 37°C (dotted lines).

(B) Convertase labeled perimCherry/cytoGFP bacteria (Green) exposed to C5b6MAC or Conv-MAC. Conditions were similar to those in (A), however C9-Cy5 was used to detect MAC pores (Red). 100 nM of C5 and C6 or C5b6 were used in combination with 100 nM C7, 20 nM C8 and 100 nM C9-Cy5. Conv + C5b6MAC and conv-MAC images were taken in separate experiments in which laser settings were adjusted to the staining intensity of C9-Cy5 to properly visualize pore distribution. Scale bars = 3 µm.

(C, D) Atomic force microscopy analysis of E. coli BL21 and MG1655 immobilized using the Poly-L-Lysine protocol. (C) Entire bacteria and high-resolution comparisons of untreated and convertase-labeled E. coli BL21 exposed to C5-C9 (Conv-MAC) for 10 minutes. Scale bars: 800 nm (left) and 30 nm (right). Height Scales: 1.01 µm (left), 8 nm (top right), 22 nm (bottom right). Width of magnification boxes (left): 42 nm, height scales: 8 nm (top left) and 13 nm (bottom left). Arrows highlight E. coli porin structures, an asterisk highlights the C5b-7 stalk. Height profiles (bottom) are shown for the white dashed lines in the images. (D) Atomic force microscopy (height and phase images) of convertase-labeled E. coli MG1655 exposed to a mixture of C5 and C6 (Conv-MAC) or preassembled C5b6 (Conv + C5b6MAC), in the presence of C7, C8, C9, FB and FD. Images were generated in the same experiment. Scale bars: 50 nm. Height scales: 15 nm. This figure and three other replicates are included in Fig. EV5B and Fig. EV5C.

Data information: (A) Data represent mean ± SD of 3 independent experiments.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 2

Figure 2. Structure and nucleotide/inhibitor binding by budding-yeast Hrr25

(A) Domain diagram of crystallized Hrr25 constructs from S. cerevisiae and C. glabrata, missing the P/Q-rich C-terminal domain. (B) Structures of S. cerevisiae Hrr251-394 K38R bound to CK1-7 (Chijiwa et al, 1989; Xu et al, 1996) and C. glabrata Hrr251-403 K38R bound to ADP, with domains colored as in (A). Bound PO4-/SO4- ions are shown as spheres. See Figure EV2A for additional C. glabrata Hrr25 structures, and Figure EV3A for SO4- ion electron density. (C) Overlay of the kinase domain of C. glabrata Hrr25 (Apo form, crystallized with 1.2 M PO4-) with rat CK1δ crystallized in the presence of tungstate ions (WO4; PDB ID 1CKJ, (Longenecker et al, 1996)) See Figure EV2B for detailed views of ion binding. (D) Stereo view of CK1-7 binding to S. cerevisiae Hrr25. Bound drug is positioned identically to a previous structure of S. pombe Cki1 bound to CK1-7 (Xu et al, 1996). (E) Stereo view of ADP binding to C. glabrata Hrr25 (formate structure; SO4- structure is equivalent, but the GxGxxG motif is disordered in that structure). All active-site residues shown in panels (D) and (E) are conserved between S. cerevisiae and C. glabrata Hrr25.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 3

Figure 3. Structure of the Hrr25 central domain.

(A) Two views of C. glabrata Hrr25 (Apo form) with domains colored as in Figure 1, showing bound PO4- ions and the previously-identified Mam1-binding residues His25 and Glu34 (Petronczki et al, 2006). (B) Two views, roughly equivalent to the views in (A), of the Hrr25 central domain, colored as a rainbow according to the schematic at top. Dotted lines in the schematic indicate disordered loops. See Figure EV1A for sequence alignment of this domain.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 4

Figure 4. Structure of the Hrr25-Mam1 complex.

(A) Domain diagram of S. cerevisiae Hrr25 (top) and Mam1 (bottom), with a schematic of Mam1 secondary structure. Zinc-coordinating cysteine and histidine residues (Cys114, His116, Cys119, and Cys152) are shown in their approximate locations in the secondary structure. See Figure EV1B for Mam1 sequence alignment. (B) Overall structure of the S. cerevisiae Hrr251-394 K38R:Mam187-191 complex, colored as in (A). View is equivalent to Figure 2B. (C) Close-up view of Mam187-191, colored as in the schematic in (A). (D) Close-up of the variant zinc knuckle motif of Mam1, with zinc anomalous difference electron density shown at 3 σ (gray) and 10 σ (red). (E) Geometry of zinc binding in Mam1. Zn-S bonds and the Zn-N bond were restrained to ~2.3 Å and ~2.0 Å, respectively, during refinement.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 5

Figure 5. Two interfaces in the Hrr25:Mam1 complex.

(A) The Hrr25:Mam1 complex with Mam1 shown as orange ribbons, and Hrr25 shown as a white surface with Mam1-binding interface #1 shown in green and #2 in magenta. (B) Stereo view of interface #1, which involves hydrophobic interactions by Mam1 L92, L97, L100, and I104, and a hydrogen-bond network between Mam1 Q101/E108/R131 and Hrr25 residues 12-15. (C) Stereo view of interface #2. Hrr25 residues H25 and E34 participate in a buried hydrogen-bond network with Mam1 residues R149 and Y158, and are surrounded by hydrophobic residues on both proteins. For clarity, Mam1 helix 4, on which R149, Y153, and Y158 are located, is not shown. (D) Pulldown assay with in vitro-translated Mam187-191 (wild-type and alanine mutants) and Hrr251-394 K38R. Mam1 mutations also disrupt Hrr25 binding when the two are co-expressed in E. coli (Figure EV6B). Bottom: Spore viability of MAM1 mutant strains (see Table EV2 for strains; strains tested are KC549, KC560, KC566, KC552, KC554, KC556, and KC558; 46-48 tetrads were dissected for each strain). “--” indicates that this mutation was not tested for spore viability.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 6

Figure 6. Mam1 binding restricts Hrr25 N-lobe mobility.

(A-C) Ribbon views of the S. cerevisiae Hrr251-394:Mam187-191 structure (form 1), CK1-7 bound S. cerevisiae Hrr251-394, and ADP-bound C. glabrata Hrr251-403 (formate condition), with Hrr25 colored according to main-chain B-factor from low (purple) to cyan (high). For each, coloring is normalized to correspond to the average of the 20 lowest (purple) or highest (cyan) main-chain B-factors in Hrr25. Mam1 (panel A) is colored in light gray. (D) Overall main-chain B-factors for the N-lobe (residues 1-85) of four structures of Hrr25 (both C. glabrata and S. cerevisiae) and the three crystallographically-unique views of the S. cerevisiae Hrr25:Mam1 complex. Values are normalized to the average main-chain B-factor for the entire Hrr25 chain (dotted line at 1.0). P-value=0.004 (student’s T-test). See Figure EV4 for residue-by-residue B-factor plots for each structure.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 7

Figure 7. Mam1 binding regulates Hrr25 kinase activity.

(A) ADP-Glo ATPase assay showing stimulation of S. cerevisiae Hrr251-394 (blue circles), Hrr251-394 K38R (red diamonds), Hrr25 full-length:Mam187-191 (black triangles), Hrr251-394:Mam187-191 (orange squares), and Hrr251-394:Mam187-191 R131A (green triangles) by bovine casein. See Figure EV6A for SDS-PAGE analysis of purified proteins. (B) Stimulation of Hrr251-394, Hrr251-394:Mam187-191, and Hrr251-394:Mam187-191 R131A by ATP, measured using an enzyme-coupled ATPase assay. (C) ADP-Glo assay showing the effect of added CK1-7 on Hrr251-394, Hrr251-394:Mam187-191, and Hrr251-394:Mam187-191 R131A. (D) Km, Vmax, and IC50 values for tested enzymes. Vmax was calculated from curves in (A) at 100 M ATP.
Structure of the S. cerevisiae Hrr25:Mam1 monopolin subcomplex reveals a novel kinase regulator (2016)
Figure 8

Figure 8. Hrr25 phosphorylates Dsn1.

(A) Phos-tag gels showing phosphorylation of purified S. cerevisiae Mtw1 complex by Hrr251-394 and Hrr251-394:Mam187-191. Asterisks denote N-terminal proteolytic cleavage products of Dsn1, with the red asterisk denoting a previously-characterized product lacking the first 171 residues (Hornung et al, 2011). (B) Model of the intact monopolin complex. Four copies of Csm1 (blue) and two of Lrs4 (green) form the core ‘V’-shaped kinetochore-binding complex (Corbett et al, 2010). Each dimer of Csm1 binds one Mam1 (orange) (Corbett & Harrison, 2012), which in turn recruits one copy of Hrr25 (blue/green). Thirty-one residues of Mam1 (192-222) separate the protein’s Hrr25-binding and Csm1-binding domains, which when fully extended could reach as far as ~120 Å.
Loss of salt tolerance during tomato domestication conferred by variation in a Na+/K+ transporter (2020)
Figure 2

Figure 2 - Identification of SlHAK20 using GWAS of root Na+/K+ ratio.

A Manhattan plot displaying the GWAS results of Na+/K+ ratio in root. The red-dashed line indicates the Bonferroni-adjusted significance threshold (P = 1.0 × 10-7). Red arrow indicates the significant SNP signal of Na+/K+ ratio associated with SlHAK20.

B The nucleotide diversity ratios between PIM and CER, and between CER and BIG on chromosome 4. The black-dashed horizontal lines indicate top 10% threshold for entire chromosome 4 (1.82 πPIM/πCER for domestication and 4.27 πCER/πBIG for improvement). The red arrows indicate the position of SlHAK20 in the sweeps. π, nucleotide diversity.

C SlHAK20-based association mapping and pairwise LD analysis. Dots represent SNPs. Indel48 is highlighted in red. The indel-2381, indel-1818 in the promoter region and five nonsynonymous variants are marked in blue. These eight variants are related to the pairwise LD diagram with dashed lines.

D Haplotypes of SlHAK20 among tomato natural variations. The distribution of Na+/K+ ratio in each haplotype group is exhibited by a box plot. The n indicates the number of accessions belonging to each haplotype. In the box plots, the middle line indicates the median, the box indicates the range of the percentiles of the total data using Turkey method, the whiskers indicate the interquartile range and the outer dots are outliers. Significant difference was determined by Student's t-test.

E Allele distribution of the SlHAK20 locus at position in PIM, CER, and BIG groups. The n indicates the accession number.
Loss of salt tolerance during tomato domestication conferred by variation in a Na+/K+ transporter (2020)
Figure 3

Figure 3 - Functional characterization of SlHAK20.

A Phylogenetic tree of the HAK/KUP/KT transporter family clade IV. The phylogenetic tree was constructed based on the amino acid sequences using the neighbor-joining method with MEGA5 in dicots and monocots.

B - D The expression pattern of SlHAK20 in tomato. The GUS activity was examined in seedling (left) and root (right) of 21-days-old SlHAK20TS-21pro:GUS transgenic tomato (B). Bars, 2 cm. Cross-sections of hypocotyl (C) and root (D) of SlHAK20TS-21pro:GUS transgenic plants. The scale bar is 400 μm in (C) and 200 μm in (D).

E Subcellular localization of SlHAK20-YFP in tomato. The SlHAK20Hap1-YFP (left) and SlHAK20Hap2-YFP (right) were localized in the plasma membrane. YFP, yellow florescence protein. FM4-64, a lipophilic styryl compound as a red fluorescent marker of plasma membrane. Bar, 20 μm.

F SlHAK20 complements the K+ uptake-defective yeast mutant assayed using a dose-response method. The K+ uptake-defective yeast mutant R5421 transformed with SlHAK20Hap1, SlHAK20Hap2, and vector (p416-GPD) were cultured in AP medium containing indicated concentrations of K+ for 24 h. The data points represent three replicates.

G Na+ uptake analysis of yeast cells expressing SlHAK20. Shown are the Na+ concentrations in the medium over the time of culture of the ANT3 yeast cells transformed with SlHAK20Hap1, SlHAK20Hap2, or empty vector. The initial NaCl concentration in the medium was 60 μM. The data are shown as the means ± SD (n = 4).

H Na+ uptake kinetic analysis of SlHAK20 in yeast cells. The curves show concentration- dependent Na+ inﬂux in yeast expressing SlHAK20Hap1 or SlHAK20Hap2 in the presence of 0, 5, 10, 20, 30, 40, 50, 60, 100, 150 or 200 μM NaCl in the medium. The data are expressed as mean ± SD (n = 3).
Loss of salt tolerance during tomato domestication conferred by variation in a Na+/K+ transporter (2020)
Figure 4

Figure 4 - Role of SlHAK20 in the salt-tolerant tomato.

A Generation of mutations in SlHAK20Hap1 by CRISPR/Cas9 using two independent single-guide RNA (sgRNA1 and sgRNA2). Sequences of SlHAK20Hap1 in wild type (TS-21), slhak20-1 and slhak20-2 mutant tomato are shown. The sgRNA-targeted sequences are underlined and the protospacer-adjacent motif (PAM) sequences are in blue. The deletion is indicated by a dashed line and the insertion is labeled with red.

B - D Na+/K+ ratio (B) and Na+ (C) and K+ (D) contents in roots of TS-21 wild type and slhak20 mutants. Data are shown as means ± SD (n = 4).

E, F Na+ (E) and K+ (F) contents in the xylem sap of TS-21 and slhak20 mutants after 50 mM NaCl treatment for the indicated days. Data represent means ± SD (n = 4).

G Shoot height of TS-21, slhak20-1, and slhak20-2. Plant height of 24-day-old TS-21, slhak20-1, and slhak20-2 were measured after growing under normal conditions for 14 days, followed by salt treatment for 14 days, and recovery for 10 days. Values are means ± SD (n = 10 plants of each genotype).

H Salt resistance of TS-21 wild type and slhak20 mutants. TS-21, slhak20-1, and slhak20-2 grown under normal growth conditions for 24 days, followed by a treatment with 200 mM NaCl for 14 days, and then recovered for 10 days. Control plants grown under normal growth conditions for 38 days. Scale bars, 5 cm.

Data information: In (B - G), P-value were determined by Student's t-test.
Loss of salt tolerance during tomato domestication conferred by variation in a Na+/K+ transporter (2020)
Figure 5

Figure 5 - Salt tolerance of slhak20 from the salt-sensitive tomato and oshak4/17 from rice.

A Generation of SlHAK20Hap2 mutations using CRISPR/Cas9 system. The sequences of SlHAK20Hap2 in wild type (TS-670), slhak20-3 and slhak20-4 mutant tomato are shown.

B Salt-tolerance assay of TS-670 and slhak20 mutant plants. TS-670, slhak20-3, and slhak20-4 grown under normal growth conditions for 23 days, and then treated with 150 mM NaCl for 5 days. Control plants grown under normal growth conditions for 28 days.

C, D Plant height and fresh weight of TS-670, slhak20-3, and slhak20-4 under normal and salt stress conditions. Shoot height (C) and fresh weight (D) of 23-day-old TS-670, slhak20-3, and slhak20-4 were quantified after growing under normal conditions for 5 days, or salt treatment with 150 mM NaCl for 5 days. Values are means ± SD (n = 10 plants of each genotype).

E Generation of oshak4/17 mutants using CRISPR/Cas9 method. Sequences of OsHAK4 and OsHAK17 in wild type (NIP), oshak4/17-1, oshak4/17-2 and oshak4/17-3 mutants are shown.

F - H Phenotype of NIP and oshak4/17 double mutant plants under normal and salt stress conditions. 20-day-old seedlings of NIP and oshak4/17 mutants were treated with 100 mM NaCl for 8 days. The pictures show representative plants (F), and the fresh weight of shoots (G) and survival rate (H) were analyzed among these genotypes. Control plants were grown under normal growth conditions for 28 days and the fresh weights of shoots were measured. Values are means ± SD (n = 12 plants of each genotype for fresh weight, four replicates for survival rate).

Data information: In (A) and (E), sgRNA-targeted sequence and a protospacer-adjacent motif (PAM) are highlighted with underline and in blue, respectively. Deletion and insertion are represented by dashed line and in red, respectively. In (C, D) and (G, H), P-value were determined by Student's t-test. Scale bar in (B) and (F), 10 cm.
Loss of salt tolerance during tomato domestication conferred by variation in a Na+/K+ transporter (2020)
Figure 6

Figure 6 - Phenotype of the Hap2 accession overexpressing SlHAK20Hap1.

A The transcript levels of SlHAK20 in TS-670 and three independent SlHAK20Hap1-YFP transgenic lines grown under normal growth conditions.

B The protein levels of SlHAK20Hap1-YFP in TS-670 and transgenic plants. ACTIN was used as loading control.

C, D Phenotypes of transgenic lines in soil under salt stress. The survival rates were obtained from at least 8 plants in four repeated experiments (C). 38-day-old plants grown in soil as control. 20-day-old TS-670 and Hap1OE-1 to -3 were treated with 175 mM NaCl for 3 weeks and recovered for 2 weeks (D).

E - G The Na+ (E) and K+ (F) contents and Na+/K+ ratio (G) in shoots and roots of TS-670 and three independent SlHAK20Hap1-YFP transgenic lines during salt stress. The 21-day-old TS-670, Hap1OE-1, Hap1OE-2 and Hap1OE-3 plants grown in 0.25× Hoagland which were then treated with 50 mM NaCl for additional 0 and 7 days.

Data information: In (A), data are shown as means ± SD (n = 6). In (C) and (E - G), data are shown as means ± SD (n = 4). P-value were determined by Student's t-test.

Expanded View Figure Legends
The transcription factor GATA4 promotes myocardial regeneration in neonatal mice (2016)
Figure 1

Figure 1: Cardiac GATA4 becomes downregulated within the first postnatal week and is necessary for myocardial regeneration.

(A) Cardiac GATA4 protein abundance analyzed by immunoblotting. GAPDH was the loading control. + denotes positive control for GATA4 from cardiomyocytes infected with a GATA4 overexpressing adenovirus.

(B) Densitometric quantification of the immunoblot shown under (A); ***p=0.0003 for P1 vs. P7 and P1 vs. P21; p<0.0001 for P1 vs. P60. The number within bars indicates the number of mice analyzed in that particular group.

(C) Immunoblot of cardiac GATA4 abundance in control (Con) and cardiomyocyte specific Gata4 knock-out mice (CM-G4-KO) at postnatal day (P) 1.

(D) Quantification of the immunoblot shown in (C); *p=0.018.

(E) Cardiac immunofluorescence staining and its quantification from Con and CM-G4-KO mice at P1. Scale bar: 25 µm; ****p<0.0001.

(F) Transversal sections of mouse hearts 7 days after cryoinjury as indicated, stained with Sirius-Red . Scale bar: 500 µm. The quantification of the left ventricular (LV) scar area (red, arrows) 7 days after injury is shown on the right; **p=0.0027.

(G) The quantification of the LV scar area of the indicated mice 21 days after cryoinjury; *p=0.0261.

(H) The quantification of the LV scar area of the indicated mice 60 days after cryoinjury; ****p<0.0001.

(I) Echocardiographic analysis of left ventricular systolic function (measured as fractional area change) in the indicated mice 7 days after cryoinjury; *p=0.033 between Con mice after sham or cryoinjury and p=0.04 between Con and CM-G4-KO mice after cryoinjury.