Journal: Journal of Sport and Health Science

Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice

doi: 10.1016/j.jshs.2025.101095

Figure Lengend Snippet: Voluntary exercise alters left ventricle gene and protein expression related to mitochondrial biogenesis and dynamics in high fat- and chow-fed mice. (A) Western blot analysis to assess purity of crude mitochondria isolation where left ventricle mitochondrial pellet expresses mitochondrial marker TOM70 and COXIV and supernatant expresses calnexin. Protein expression of (B) LCLAT1 and (C) MFN2 in left ventricle-isolated crude mitochondria and (D) LCLAT1, (E) PGC-1α, and (F) MFN2 in left ventricle tissue from male VET or sedentary mice fed an HFD or chow diet. Left ventricle mRNA expression of (G) OPA1 and (H) DRP1 in male VET or sedentary mice fed an HFD or chow diet. Values were calculated relative to 18S housekeeper. Calnexin and β-actin were used as internal controls of protein loading in left ventricle samples and COXIV as internal control of protein loading in crude mitochondria samples. Analysis was performed using two-way analysis of variance with Tukey’s post hoc test for multiple comparisons. Data are expressed as mean ± standard error of the mean. (B–C) n : 5–6 per group. (D–F) n = 7 per group. (G and H) n = 9 per group. CE = chow exercise; CS = chow sedentary; COXIV = cytochrome c oxidase subunit 4; DRP1 = dynamin-related protein 1; HE = high fat diet exercise; HFD = high fat diet; HS = high fat diet sedentary; LCLAT1 = lysocardiolipin acyltransferase 1; MFN2 = mitofusin-2; OPA1 = optic atrophy 1; PGC-1α = peroxisome proliferator-activated receptor gamma coactivator-1α; TOM70 = translocase of outer mitochondria membrane 70; VET = voluntary exercise training.

Article Snippet: Immunoblotting was performed with antibodies against lysocardiolipin acyltransferase 1(LCLAT1, PA5-25627; Thermo Fisher Scientific), PGC-1α (ab191838; Abcam, Cambridge, UK), MFN2 (PA5-118059; Thermo Fisher Scientific), ANP (sc-18811; Santa Cruz, Dallas, TX, USA), tumor necrosis factor alpha (TNFα, 3707; Cell Signaling, Danvers, MA, USA), phosphorylated adenosine monophosphate-activated protein kinase (AMPK, 2531; Cell Signaling), AMPKα (2532; Cell Signaling), Perilipin 5 (PA1-46215; Thermo Fisher Scientific), translocase of outer mitochondria membrane 70 (TOM70, 65675; Cell Signaling), cytochrome c oxidase subunit 4 (COXIV, 4844; Cell Signaling), and calnexin (208880; Merck, Rahway, NJ, USA).

Techniques: Expressing, Western Blot, Isolation, Marker, Control, Membrane

Journal: iScience

Article Title: 3D EM uncovers mitochondrial network remodeling in residual triple negative breast cancer after conventional chemotherapy treatments

doi: 10.1016/j.isci.2026.115865

Figure Lengend Snippet: PIM001-P PDX tumors persisted following conventional chemotherapy treatments (A) Schematic of the experimental approach used in the study. Images were adapted from BioRender. (B) Tumor volumes were monitored biweekly following the administration of chemotherapies to mice bearing orthotopic PIM001-P tumors ( n = 6 mice/group). Treatment groups were treatment-naïve (TN), Adriamycin + cyclophosphamide (AC), docetaxel + carboplatin (DTX+CRB), docetaxel (DTX), and carboplatin (CRB). Arrows at the top indicate when chemotherapy was administered. Asterisks above the x axis indicate early euthanasia due to animal health concerns. Residual tumors were harvested, as noted by arrows. Error bars represent the standard error of the mean. (C) Residual tumors were harvested, processed for FFPE, and analyzed by H&E staining and IHC for Ki67, Ku80, and human-specific mitochondria. Scale bars are 50 μm.

Article Snippet: Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (f) 3D volume, (g) 3D area, (h), and perimeter.

Techniques: Staining

Journal: iScience

Article Title: 3D EM uncovers mitochondrial network remodeling in residual triple negative breast cancer after conventional chemotherapy treatments

doi: 10.1016/j.isci.2026.115865

Figure Lengend Snippet: Single-agent chemotherapy treatments in PIM001-P significantly increased mitochondrial size in residual tumors relative to TN (A–E) Representative orthoslices. (a’-e’) Representative orthoslices with overlays of mitochondria segmentations. (a’’-e’’) Representative mitochondria segmentations for each treatment group: treatment-naïve (TN), adriamycin + cyclophosphamide (AC), docetaxel + carboplatin (DTX+CRB), docetaxel (DTX), and carboplatin (CRB). The scale bars are 3 μm. Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (f) 3D volume, (g) 3D area, (h), and perimeter. Adjusted p -values (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001) are shown on box and whisker plots as determined using the Mann-Whitney U test followed by the Holm method.

Article Snippet: Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (f) 3D volume, (g) 3D area, (h), and perimeter.

Techniques: Software, Whisker Assay, MANN-WHITNEY

Journal: iScience

Article Title: 3D EM uncovers mitochondrial network remodeling in residual triple negative breast cancer after conventional chemotherapy treatments

doi: 10.1016/j.isci.2026.115865

Figure Lengend Snippet: Single-agent chemotherapy treated residual tumors’ mitochondria are more branched and less spherical relative to TN in PIM001-P (A) Representative longitudinal and transverse views of the segmented mitochondria for each treatment group: treatment-naïve (TN), adriamycin + cyclophosphamide (AC), docetaxel + carboplatin (DTX+CRB), docetaxel (DTX), and carboplatin (CRB). The scale bars are 3 μm. Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (B) sphericity (C) mitochondrial complex index (MCI), along with their respective method of measurements shown as a cartoon above the bar graphs. Adjusted p -values (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001) are shown on box and whisker plots as determined using the Mann-Whitney U test followed by the Holm method.

Article Snippet: Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (f) 3D volume, (g) 3D area, (h), and perimeter.

Techniques: Software, Whisker Assay, MANN-WHITNEY

Journal: iScience

Article Title: 3D EM uncovers mitochondrial network remodeling in residual triple negative breast cancer after conventional chemotherapy treatments

doi: 10.1016/j.isci.2026.115865

Figure Lengend Snippet: PIM001-P mito-otyping of the diverse mitochondrial structures within each treatment group (A) Representative mitochondria structures from the bottom, middle, and top 10% of the 3D volume for tumors are displayed. Scale bars are 1 μm.

Article Snippet: Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (f) 3D volume, (g) 3D area, (h), and perimeter.

Techniques:

Journal: iScience

Article Title: 3D EM uncovers mitochondrial network remodeling in residual triple negative breast cancer after conventional chemotherapy treatments

doi: 10.1016/j.isci.2026.115865

Figure Lengend Snippet: The frequency of MLCs increases, while the sizes of the contacts decrease following chemotherapy Quantification of how chemotherapy remodels mitochondrial-lipid droplet contacts (MLCs) for each WHIM14 treatment group: treatment-naïve (TN), docetaxel + carboplatin (DTX+CRB), docetaxel (DTX), and carboplatin (CRB). (a-d’’) 3D rendering of the mitochondria in purple, lipid droplets in yellow, and their interaction in red. The scale bars are 2 μm. The 3D renders show that TN exhibits relatively few but broad, sheet-like interfaces, whereas all treatment groups display numerous, smaller, punctate contacts scattered across organelles. Consistent with this visual shift, bar plots demonstrate that the fraction of MLCs among the total number of segmented mitochondria (E; % MLC per 500 mitochondria) is higher after treatment. In contrast, per-contact surface area (F) and volume (G) are largest in TN and significantly reduced in all treated conditions, indicating that individual interfaces between the mitochondria and LDs shrink following chemotherapy. This is also displayed in the bar plots, showing the total sum of either surface area (H) or volume of the MLC versus the total sum of each metric for the segmented mitochondria.

Article Snippet: Mitochondrial measurements were calculated using Amira software for 350 segmented mitochondria (represented as dots) from each tumor for (f) 3D volume, (g) 3D area, (h), and perimeter.

Techniques: