EMBO/sd-character-level-ner huggingface dataset:
a) HeLa cells transfected for 24 h with Flag-tagged wild-type Atg16L1 or the Atg16L1 deletion mutants 2–275 or 232–607 Atg16L1, together with GFP–Atg5 and CFP–LC3, were fixed and immunostained with anti-Flag antibody before confocal analysis.figf6(a) HeLa cells transfected for 24 h with Flag-tagged wild-type Atg16L1 or the Atg16L1 deletion mutants 2-275 or 232-607 Atg16L1, together with GFP-Atg5 and CFP-LC3, were fixed and immunostained with anti-Flag antibody before confocal analysis. Co-localization of wild-type or 2-275 Atg16L1 (red) with GFP-Atg5 (green) is marked by arrows in the insets. Scale bars, 10 μm. (b) HeLa cells transfected for 24 h with GFP-Atg16L1 were incubated for 15 min with horseradish peroxidase (HRP)-conjugated cholera toxin subunit B at 4 °C. The cells were then incubated for 10 min at 37 °C, after which they were processed for double immunogold labelling with anti-HRP antibodies (15-nm gold particles) and anti-GFP antibodies (10-nm gold particles) for electron microscopy. PM, plasma membrane. Scale bar, 100 nm. Co-localization can be seen in boxed areas. n = 49 fields. All error bars represent s.e.m.
Figure 3A Immunoprecipitation assay was performed using anti-Flag antibody in MM-231-shLSD1-WT/R838A cells. The interaction of LSD1 and DUBs were detected by immunoblotting with indicated antibodies. B MDA-MB-231 cells were treated with the indicated amounts of P5091 for 48 h, LSD1 protein levels were assessed by immunoblotting. C MDA-MB-231 cells were treated with P5091 (10 μM) for 48 h and MG132 for 10 h. The LSD1 ubiquitination level was measured by immunoblotting after immunoprecipitated with anti-LSD1 antibody. D HEK-293T cells were transfected with USP7 WT or USP7 C223S, then treated with MG132 for 10 h. Cell lysates were immunoprecipitated using anti-LSD1 antibody followed by immunoblotting analysis. E MDA-MB-231 cells overexpressing Flag-tagged LSD1 WT/R838A after knockdown of endogenous LSD1 and then treated with or without P5091 for 48 h and MG132 for 10 h. Cell lysates were immunoprecipitated by anti-Flag antibody and then analysed by immunoblotting. F MDA-MB-231 cells were transfected with control or CARM1 shRNAs before treated with MG132 for 10 h. And cell lysates were immunoprecipitated with anti-LSD1 then subjected to immunoblotting. G MDA-MB-231 cells were treated with or without MS049 for 48 h and MG132 for 10 h, Cell lysates were immunoprecipitated using anti-LSD1 antibody and then analysed by immunoblotting. H HEK-293T cells were transfected with USP7 WT or USP7 C223S, then treated with or without MS049 for 48 h and MG132 for 10 h. Cell lysates were immunoprecipitated with anti-LSD1 antibody followed by immunoblotting analysis.
Figure 1A. Representative FACS plots of freshly isolated epidermal cells (Epi d0) and day 14 (d14) cultures of epidermal cells in different growth conditions and media. KGM 2D: 2D in basal KGM; 3C: 3D-Matrigel in KGM + Y27632 + VEGF-A + FGF-2. B. CD34+α6+ cells were quantified by flow cytometry from d14 as in (A) (mean ±SD; n=3-5; *p ≤ 0.05; ***p ≤ 0.001, Kruskal-Wallis/Dunn's post test). 3C 2D: 2D in KGM + Y27632 + VEGF-A + FGF-2; Y: 3D-Matrigel in KGM + Y27632; YV: Y + VEGF-A; YF: Y + FGF-2; 3C: Y + VEGF-A + FGF-2. C. Absolute numbers of CD34+α6+ cells from Epi d0 and d14 cultures as in (A-B). Fold enrichment over Epi d0 is shown (mean ±SEM, n=3-5; *p ≤ 0.05, Mann-Whitney U test). D. Epidermal cells grown for 14 days in the indicated conditions. 3C: 3D-Matrigel in KGM + Y27632 + VEGF-A + FGF-2. 3C 2D: 2D in 3C medium. Scale bars 30µm. E. Immunofluorescence analysis of 3C cultures showing the expression of the HFSC markers CD34 and SOX9 in a subset of cells. See also Appendix Appendix figure S1.
Figure 3A, Summary of chimera assays in which one single Td-Tomato fluorescent-labeled human iPSC, either control or transiently transfected with miR-203-expressing vectors (miiPSCs) was injected into 8C-stage mouse embryos and their contribution analyzed 48-60 h later. P values are indicated in the Figure (Student´s t-test). B, Representative images showing the contribution of human control iPSCs or miiPSCs cells to mouse blastocysts. These structures were co-immunostained with (human) anti-OCT4 and anti-HuNu antibodies. Td-Tomato, red; Scale bars, 50 μm. C, Interspecies chimera assays in which the 8C-stage mouse embryos indicated in (A, B), injected with one single human iPSCs and cultured in vitro for 48 h to reach the blastocyst stage, were then transfected to 2.5 days post coitum pseudo-pregnant females. The post-implantation mouse conceptuses were dissected at the E9.5 developmental stage and analyzed by immunofluorescence. Representative images showing the integration of control human iPSCs (top) or miiPSCs (bottom) into mouse E9.5 embryos. Anti-human HuNu antibody was co-stained with Td-Tomato direct fluorescence to detect human cells in E9.5 mouse embryos. Scale bars, 20 μm. D, E, Quantification of the Td-Tomato-(D) or HuNu-(E) positive area in 40-50 different cryosections of these embryos randomly and blindly selected for IF analysis. Data are mean ± s.e.m. **P<0.01; ***P< 0.001 (Student´s t-test).
Figure 5A, B RT-qPCR analysis of Ifnb1 mRNA level in the WT and lincRNA-EPS-/- iBMMs infected with VSV (MOI 0.1) for 6 h (A), ELISA analysis of supernatant IFN-β protein after infecting for 8 h (B). C, D Mx1 (C) and Oas2 (D) mRNA level were measured in the control (EV) and lincRNA-EPS-rescued (lincRNA-EPS) iBMMs after infecting with VSV with different titer for 6 h. E LincRNA-EPS-stably overexpressed iBMMs (lincRNA-EPS) and corresponding control group (EV) were infected with VSV for different titer (MOI 0.1 and MOI 0.5), Mx1 and Oas2 mRNA level were measured by RT-qPCR after 6 h. F The basal level of ISGs and proinflammatory genes in the WT and lincRNA-EPS-/- BMDMs (Datasets from the ArrayExpress database under the accession number E-MTAB-4088) were listed by heatmap. G ChIP assays using anti-H3K4me3 antibody was carried out in WT and lincRNA-EPS-/- iBMMs with or without polyI:C (1 μg/mL) transfection for 4 h, Mx1 and Oas2 enrichment relative to input were measured by qPCR. H, I WT and lincRNA-EPS-/- iBMMs were infected with VSV (MOI 0.1) for 6 h or 10 h (H) or transfected with polyI:C (1 μg/mL) for 4 h or 8 h (I), phosphorylation and total protein expression were analyzed by Western blot with α-tubulin and β-Actin as loading controls.
PRMT5 regulates splicing in ground state ESCsSchematic model of ESC derivation.Figure 5A Schematic model of ESC derivation. Recombination is induced with addition of 4‐OHT in culture for 48 h. B Western blot of iPHet and iPKO ESCs 5 days after treatment with 4‐OHT. Nu, nuclear fraction. Cyto, cytoplasmic fraction. ACTIN and H2A are used for loading control of nuclear and cytoplasmic fraction, respectively. C IF for Ki67 (green) in iPHet and iPKO ESCs 5 days after treatment with 4‐OHT. White arrows mark Ki67‐positive ESCs. Red arrows mark Ki67‐negative ESCs. Scale bar, 20 μm. D Growth curve of iPHet and iPKO ESCs at days 2, 3 and 5 after 4‐OHT treatment. n = 2 replicates. The original seeding on day 0 is 50,000 (50K) cells per well of a 6‐well plate. E Western blot for SYM10 antibody of iPHet and iPKO ESCcytoplasmic lysates 5 days after treatment with 4‐OHT. Cyto, cytoplasmic fraction. ACTIN is used as a loading control. The absence of PRMT5 is shown in iPKO ESCs. F Pair‐wise analysis of differential splicing events in each iPKO replicate relative to the paired iPHet control. A3SS, alternative 3′ splice site; A5SS, alternative 5′ splice site; MXE, mutual exclusive exons; RI, retained introns; SE, skipped exons. G Venn diagram of pair‐wise analysis of MATS, showing overlapping differential splicing events among replicates. H Representative GO terms for common differentially spliced genes identified in three out of four replicate pairs. I (Reverse transcript) RT-PCR validation of aberrant splicing events in Mdm4 and Ehmt2. SE, skipped exon. PCR is performed among at least three replicates for each locus. Electrophoresis image was quantified using ImageJ software. **P = 0.0046, ***P = 0.0006. J Representative flow plot of sorting cKIT+/SSEA1+PGCs (red box) at E10.5 using FACS (shown is an embryo with genotype Prmt5fl/fl;creER with 4‐OHT injection at E6.5). K Sorted PGC numbers among control iPHet (Prmt5fl/+;creER) and mutant iPKO (Prmt5fl/fl;creER). *P = 0.0486. L PCR validation of aberrant splicing events in Mdm4 and Ehmt2 using cDNA reverse‐transcribed and amplified from FACS‐sorted PGCs (individual embryos) using the NuGEN Ovation RNA Amp System V2. SE, skipped exon. PCR was performed among one control (Prmt5fl/+;creER) and two mutants (Prmt5fl/fl;creER).
Figure 5Volcano plots showing the enrichment of RBPs which bind to REL-WT relative to REL-OPT transcripts (A) Data information: vertical dotted lines indicate a 1.5-fold enrichment while horizontal dashed lines indicate the p-value cut-off of 0.05. Points shaded in blue indicate RBPs which have a differential fold change of more than 1.5 and p<0.05. Volcano plots showing the enrichment of RBPs which bind to , IL6-DE relative to IL6-WT transcripts (B) Data information: vertical dotted lines indicate a 1.5-fold enrichment while horizontal dashed lines indicate the p-value cut-off of 0.05. Points shaded in blue indicate RBPs which have a differential fold change of more than 1.5 and p<0.05. Volcano plots showing the enrichment of RBPs which bind to IL6-WT relative to IL6-OPT transcripts (C). Data information: , vertical dotted lines indicate a 1.5-fold enrichment while horizontal dashed lines indicate the p-value cut-off of 0.05. Points shaded in blue indicate RBPs which have a differential fold change of more than 1.5 and p<0.05.
Distribution of BREX systems across the phylogenetic tree of bacteria and archaeaShown is the common tree of bacteria and archaea as represented in the NCBI Taxonomy resource ().Figure 6Shown is the common tree of bacteria and archaea as represented in the NCBI Taxonomy resource (). Organisms in which a BREX system exists are colored; color code follows the BREX subtype coloring from Fig . Extensive horizontal transfer is observed by the lack of coherence between the species tree and the PglZ phylogeny.
Figure 2A) Oxygen consumption was analyzed in permeabilized muscle fibers from tibialis muscle from young and old control and Mfn2KO mice (n=6 mice per group). B) Mitochondrial mass was evaluated by mtDNA copy number in gastrocnemius muscle (n=6 per group). Data were expressed as a fold change compared to young control mice. C and D) Representative images of NADH-TRstaining (C) in muscle sections from young and old control and Mfn2KO mice (n=3 mice per group). C and D) Representative images of SDHstaining (D) in muscle sections from young and old control and Mfn2KO mice (n=3 mice per group). E) Levels of H2O2 measured in gastrocnemius muscle from young and old control and Mfn2KO mice (n=6 mice per group). F) Protein carbonylation was measured in gastrocnemius muscle from young and old control and Mfn2KO mice (n=6 mice per group). Data were expressed as a fold change compared to young control mice G) Representative TEM images of longitudinal sections of quadriceps muscle from young and old control and Mfn2KO mice and quantification of mitochondrial area and density (n=3 mice per group) (Scale bar: 0.4 µm). H) Representative TEM images of aberrant mitochondria highly present in Mfn2KO old mice. Data represent mean ± SEM. #p<0.05 Mfn2KO vs. control mice, *p<0.05 old vs. young mice.
Macrophages treated with MDP after M.Figure 6Macrophages treated with MDP after M. tuberculosis infection develop autophagy vacuoles. Alveolar macrophages were infected with M. tuberculosis H37Rv at an MOI of 5 for 1 h. Nonphagocytosed bacteria were washed away, and the macrophages were incubated for an additional hour. The cells were then treated with 10 μg/mL of MDP for 24 h. Autophagy indicators (arrowheads) such as (A) vesicles containing organelles (TEM magnification ×20,000) and (B) onion skin‐like lamellar multivesicular bodies (TEM magnification ×16,000) were identified by ultrastructural analysis.. (C, D) Autophagosomes containing mycobacteria (Mtb) are indicated with arrows (C, TEM magnification ×40,000; D, TEM magnification ×12,000). Micrographs were obtained from one experiment. Bar represents 300 nm.
Figure 1A. Representative images of immunofluorescence of α-smooth muscle actin (α-SMA), pan Cytokeratin, Pan-hMENA, hMENA11a and E-cadherin expression in CAFs and autologous cancer cells (Ep-PDAC) obtained from enzymatically digested primary PDAC tissue of patient #36. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar: 20 μm. B. Representative immunoblot (top) of hMENA/hMENA∆v6 expression levels (detected by Pan-hMENA mAb and by the specific anti-hMENAΔv6 antibody) in normal fibroblasts derived from transplant donor (P-NF), pancreatic serous cystadenoma #71 and cancer associated fibroblasts (n=10) obtained from primary PDAC tissues. Densitometry quantified data (bottom) of hMENA∆v6 expression. Quantification of P#110 is relative to the sample shown in the WB on the left. C. Representative immunoblot (top) of hMENA/hMENA∆v6 expression levels (detected by Pan-hMENA mAb and by the specific anti-hMENAΔv6 antibody) in normal lung fibroblasts (L-NF), cancer associated fibroblasts obtained from NSCLC tissues (n=4) and normal dermal fibroblasts (HNF). Densitometry quantified data (bottom) of hMENA∆v6 expression.
The role of p38IP in p38α‐dependent inhibition of autophagy.Figure 5(A) 293/GFP-LC3 cells were pre‐treated with 10 μM anisomycin for 30 min before incubation in either full medium or EBSS. Cell lysates were analysed by SDS-PAGE for GFP-LC3 lipidation using an anti‐LC3 antibody. The membrane was also probed with anti‐β tubulin. The GFP-LC3 lipidation was quantified as the amount of GFP-LC3II/GFP-LC3I (data are represented as mean±s.e.m. of triplicates, P=0.0091, Student's t‐test). (B) HEK293A cells labelled with [14C]valine, as described in Material and Methods section, were pretreated with 10 μM anisomycin for 30 min before incubation in either full medium, EBSS, or EBSS with leupeptin for 2 h. Cells were then analysed for long‐lived protein degradation (data are represented as mean±s.e.m. of triplicates, representative of two experiments, EBSS untreated versus anisomycin treated (P=0.0363); EBSS untreated versus leupeptin treated (P=0.0001), Student's t‐test). (C) HEK293 cells were transfected with control siRNA or siRNA for p38IP. After 24‐h incubation, control or p38IP‐depleted cells were pre‐treated with anisomycin, followed by a 2‐h incubation with full medium, full medium with leupeptin, EBSS, or EBSS with leupeptin. Cell lysates were analysed with anti‐p38α or anti‐LC3 antibodies. LC3II/LC3I levels were quantified and presented normalized to control, untreated cells incubated in EBSS. Data are representative of two experiments. (D) HEK293A cells transiently transfected with HA‐p38IP and RFP‐mAtg9 were treated with anisomycin for 30 min and mAtg9 was immunoprecipitated. 5% of the lysates or the immunoprecipitates were probed with anti‐HA and anti‐mAtg9 antibodies (data is representative of 3 experiments, *P=0.0332).
Figure 6(A) Left panel shows the individual channels and overlaid images of confocal live cell imaging of HeLa cells overexpressing GFP-Bax (green) and mito-dsRed (magenta) at 0 and 85 minutes after apoptosis induction with STS 1µM. Right panel shows zoomed overlaid images of HeLa cells following GFP-Bax translocation (green) and mitochondrial fragmentation (magenta) during apoptosis induction with STS 1µM. Scale bars, 10 µm (left) and 2 µm (right)(B) Reconstructed dual-color superresolution image of GFP-Bax overexpressing HeLa cells. GFP-Bax was imaged by immunosstaining with AF647-anti-GFP-nanobodies and mitochondria were immunostained with CoxIV (primary) and cf680 (secondary) antibodies. Image was acquired 3h after STS treatment. Left panel shows the cell overview (scale bar, 5 µm), right panel up shows the overview inset (scale bar, 500nm) and right panel down shows a non-random linear GFP-Bax assembly at mitochondria (scale bar, 100nm).(C) Reconstructed dual-color superresolution image of HeLa cells co-overexpressing mMaple-mito and GFP-Bax and stained with AF647-anti-GFP-nanobodies. Image was acquired 3h after STS treatment. Lower panel shows enlarged insets corresponding to the white rectangles in the upper panel. Scale bars, 1 µm (overview) and 100 nm (insets)
after fixation with either PFA or glyoxal.Figure 5 Figure 5 Comparison of immunostaining NUP160 after fixation with either PFA or glyoxal. HeLa cells were stained for the nucleoporin complex protein NUP160 after fixation with either PFA, glyoxal pH 4 or glyoxal pH 5. Fluorescence intensities (fold over background) were compared and are shown in the graph. The quantification of fluorescence signals shows that glyoxal pH 4 fixation allows for significantly brighter stainings. N = 73 - 156 cells per condition analyzed. Scale bar = 10 µm. *** p < 0.001
Spatial distribution of neuronal subtypes in the hippocampus.Figure 4a. (i) Cell subtype map of CA1 neurons in the hippocampus as annotated by JSTA. Scale bar is 500 micron. Distribution of CA1 subtypes in the hippocampus, computed by projecting cell centers to the lateral to medial axis. CA1 neuronal subtypes show a non-uniform distribution across the whole CA1 region. (ii) Smoothed histogram highlighting the density of CA1 subtypes across the CA1 region. b. (i) Cell subtype map of CA3 neurons in the hippocampus as annotated by JSTA. Distribution of CA3 subtypes in the hippocampus, computed by projecting the cell centers to the lateral to medial axis. CA3 neuronal subtypes show a non-uniform distribution across the whole CA3 region. (ii) Smoothed histogram highlighting the density of CA3 subtypes across the CA3 region.
Silencing ATG-5 sensitized chemoresistant cells to drug treatment.Figure 4(A) The mRNA level of ATG-5 was detected by real time PCR after treatment with siRNAs. GAPDH was used as internal controls. (B) The protein level of ATG-5 was detected by western blot after treatment with siRNAs. β -actin was used as internal controls. (C) The proliferation ability was tested using MTT assay 48 hours or 72 hours after different treatment.
FGF7 suppresses poly(I:C)-induced IFN and ISG expression (A) Serum-starved HaCaT cells were treated for 24 h with poly(I:C) (5 μg/ml) in the presence or absence of FGF7 (10 ng/ml).Figure 4(A) Serum-starved HaCaT cells were treated for 24 h with poly(I:C) (5 μg/ml) in the presence or absence of FGF7 (10 ng/ml). RNA samples were analyzed by qRT-PCR for IFNL1, IFNB, RSAD2, IRF1, IRF7, CGAS, TLR3 or DDX58, relative to RPLP0. (B) Serum-starved HaCaT cells were treated for 18 h with poly(I:C) (5 μg/ml) in the presence or absence of FGF7 (10 ng/ml). Protein lysates were analyzed by Western blot for total and pSTAT1 (Y701 and S727), total and pSTAT2 (Y690), IRF1, IRF3, pIRF3 (S396), IRF9, RIG-1, RSAD2 and GAPDH.
Figure 2Olfm4-positive intestinal stem cells (indicated by arrows) in small intestine sections (A). Quantification of intestinal stem cell (Olfm4+) number per 100 μm (n=4-5 mice per group) of small intestine sections (B). Representative images of small intestine crypt culture (day 2). Arrows mark the surviving cysts (C). Quantification of numbers of cysts per culture well (D)(n=4-5 mice/genotype). In vitro crypt culture (day 8) from small intestine crypts isolated from mice with indicated genotypes. Arrows mark the crypt-like structure (E). The number of organoids per culture wells (n=4-5 mice per group). Two hundred crypts are seeded per well (F). Representative images of TUNEL staining in small intestine sections. Arrows indicate TUNEL positive cells (G). Quantification of TUNEL positive cells per crypt (n=5 mice per group) (H). Representative images of phosphorylated histone 3 (pH3) antibody staining in mouse small intestine. Arrows indicate pH3 positive cells (I). Quantification of pH3 positive crypt cells per 100 μm of small intestines (n=4-6 mice per group) (J).
EGFR is required and sufficient in the lamina glia to maintain glia integrity.Fig 4(A) repots>Shits1+H2B-RFP, (B) repots>Shits1+Egfrλtop and (C) repots>Shits1+RlSem incubated for 12 days at 28°C. (D) repots>DERDN for 7 days at 28°C. (E) Epithelial glia MARCM clone and (F) Egfrco mutant glia MARCM clone (labeled with Tomato, green). The penetrance is indicated as the number of samples with vacuoles over the total number of samples. DAPI: nuclei (white in A-F). (G) Percentage of vacuole area in the lamina neuropile of (A-D). The P-values were calculated using one-way ANOVA with Dunnett's post-test. (H) Percentage of vacuole area in the lamina of repots>Shits1 flies in the indicated genetic background. The adults were shifted to 28°C for 12 days. The P-values were calculated using one-way ANOVA with Tukey's post-test. Scale bar: 20 μm.
Autophagy was involved in the drug resistant of DC cells.Figure 5(A) The autophagy was detected by immunofluorescence assay of LC3B in the cells 48 hours after different treatment. (B) The protein levels of LC3A and LC3B were tested by western blot. β-actin was used as internal controls.
TLR8 ligands induce autophagy in human macrophages.Figure 1Macrophages were treated for 24 h with CL097, ssRNA40, ssRNA41 or rapamycin (Rapa). (A) Immunoblot of co-immunoprecipitation of cell lysates with BECN1 specific antibody using antibody to PIK3C3 and BECN1 with antibody to β-actin as loading control. (B) Cells were incubated with 10 µg/mL pepstatin A for 4 h prior to lysis. Immunoblots of LC3B isoforms using antibody to LC3B or β-actin. (C) Cells were fixed and permeabilized then stained with antibody to LC3B (green) and Hoechst 33342 (blue). Scale bars indicate 10 µm. Left, representative fluorescence microscopy images. When puncta are not present, LC3B staining gives rise to a diffuse cytoplasmic pattern. Right, percentage of cells with LC3B puncta. Bar charts are presented as the means ± s.e.m., n = 6. (D) Immunoblots of SQSTM1 using antibody to SQSTM1 or β-actin. * p<0.05.
a) NRK cells were transfected with nonspecific (NS)- or Cltc (clathrin heavy chain)-RNAi and starved for 0, 4, 8 or 12 h.figf8(a) NRK cells were transfected with nonspecific (NS)- or Cltc (clathrin heavy chain)-RNAi and starved for 0, 4, 8 or 12 h. Scale bar, 5 mm. Cells were stained with antibodies against LC3. (b) Cells from a were assessed for LC3 puncta in a blind fashion after starvation and quantified. n = 100 cells from three independent experiments. Error bars indicate the s.d.
Mitochondrial clearance is impaired in Nix−/− reticulocytes transduced with Nix‐W35A.Figure 4(A) Flow cytometry of reticulocyte‐enriched blood, cultured in vitro for 3 days, stained with MTR. The blood was taken from mice transplanted with Nix−/− bone marrow and reconstituted with wt‐Nix and Nix‐W35A viruses. Reticulocytes were induced by phenylhydrazine treatment. The viruses also express GFP. (B) Mitochondrial clearance in the GFP‐positive fraction of the reticulocyte‐enriched blood, as described in panel (A). Mice were reconstituted with wt‐Nix, LIR‐W35A or empty vector viruses. Data from Nix−/− and Nix+/+ mice are shown for comparison. (C) Anti‐Flag, anti‐GFP and anti‐β‐actin western blots of fetal liver cells transduced with Flag‐Nix and Flag‐W35A‐Nix viruses, normalized to total protein. GFP, green fluorescent protein; LIR, LC3‐interacting region; MTR, MitoTracker Red; wt, wild type.