Review




Structured Review

MitoSciences total oxphos rodent antibody cocktail ms604
Total Oxphos Rodent Antibody Cocktail Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pm40616270-68-35-39
Average 90 stars, based on 1 article reviews
total oxphos rodent antibody cocktail ms604 - by Bioz Stars, 2026-09
90/100 stars

Images

Related Articles

Saline:

Article Title: The proteoglycan decorin does not influence adiposity, glucose tolerance, or aerobic exercise capacity in mice.
Article Snippet: For total protein measurements the membranes were either stained with Ponceau S, or visualized with TGX stain- free technology (Bio- Rad, Hercules, CA, USA) for fluorescent detection of proteins. .. The membranes were then blocked in Tris buffered saline containing 0.1% Tween- 20 and 5% BSA, and incubated over night with primary antibodies in 2.5% BSA in TBS- T. The following antibodies were used: Total OXPHOS rodent antibody cocktail (MitoScience LLC Cat# MS604, RRID:AB_2629281), glycogen synthase (Cell Signaling Technology Cat# 3886, RRID:AB_2116392), phospho- glycogen synthase (Cell Signaling Technology Cat# 3891, RRID:AB_2116390), Ppargc1a (Santa Cruz Biotechnology Cat# sc- 13067, RRID:AB_2166218), Tnf (Cell Signaling Technology Cat# 3707, RRID:AB_2240625), PI3 Kinase p85 (Millipore Cat# 06- 497, RRID:AB_310141), Stat- 3 (Cell Signaling Technology Cat# 4904, RRID:AB_331269), and phopho- Stat3 ((Cell Signaling Technology Cat# 9167, RRID:AB_561284)). .. For detection we used HRP- conjugated secondary antibodies (Jackson ImmunoResearch Labs Cat# 111- 035- 144/RRID:AB_2307391 and Cat# 115- 035- 146/ RRID:AB_2307392), ECL substrate (Cat# 34076, Thermo Scientific, Rockford, IL, USA) and the ChemiDocTM Touch Imaging System (Bio- Rad, Hercules, CA, USA).

Article Title: Time dynamics of elevated glucose and beta-hydroxybutyrate on beta cell mitochondrial metabolism.
Article Snippet: .. Membranes were blocked for 2 hours at room temperature with fat-free milk (5% w/v), Tween 20 (0.1%) in Tris-buffered saline, pH 7.6, and then incubated overnight at 4°C with primary antibodies for oxidative phosphorylation complexes MS604, (Mitosciences, USA). ..

Incubation:

Article Title: The proteoglycan decorin does not influence adiposity, glucose tolerance, or aerobic exercise capacity in mice.
Article Snippet: For total protein measurements the membranes were either stained with Ponceau S, or visualized with TGX stain- free technology (Bio- Rad, Hercules, CA, USA) for fluorescent detection of proteins. .. The membranes were then blocked in Tris buffered saline containing 0.1% Tween- 20 and 5% BSA, and incubated over night with primary antibodies in 2.5% BSA in TBS- T. The following antibodies were used: Total OXPHOS rodent antibody cocktail (MitoScience LLC Cat# MS604, RRID:AB_2629281), glycogen synthase (Cell Signaling Technology Cat# 3886, RRID:AB_2116392), phospho- glycogen synthase (Cell Signaling Technology Cat# 3891, RRID:AB_2116390), Ppargc1a (Santa Cruz Biotechnology Cat# sc- 13067, RRID:AB_2166218), Tnf (Cell Signaling Technology Cat# 3707, RRID:AB_2240625), PI3 Kinase p85 (Millipore Cat# 06- 497, RRID:AB_310141), Stat- 3 (Cell Signaling Technology Cat# 4904, RRID:AB_331269), and phopho- Stat3 ((Cell Signaling Technology Cat# 9167, RRID:AB_561284)). .. For detection we used HRP- conjugated secondary antibodies (Jackson ImmunoResearch Labs Cat# 111- 035- 144/RRID:AB_2307391 and Cat# 115- 035- 146/ RRID:AB_2307392), ECL substrate (Cat# 34076, Thermo Scientific, Rockford, IL, USA) and the ChemiDocTM Touch Imaging System (Bio- Rad, Hercules, CA, USA).

Article Title: Time dynamics of elevated glucose and beta-hydroxybutyrate on beta cell mitochondrial metabolism.
Article Snippet: .. Membranes were blocked for 2 hours at room temperature with fat-free milk (5% w/v), Tween 20 (0.1%) in Tris-buffered saline, pH 7.6, and then incubated overnight at 4°C with primary antibodies for oxidative phosphorylation complexes MS604, (Mitosciences, USA). ..

Phospho-proteomics:

Article Title: Time dynamics of elevated glucose and beta-hydroxybutyrate on beta cell mitochondrial metabolism.
Article Snippet: .. Membranes were blocked for 2 hours at room temperature with fat-free milk (5% w/v), Tween 20 (0.1%) in Tris-buffered saline, pH 7.6, and then incubated overnight at 4°C with primary antibodies for oxidative phosphorylation complexes MS604, (Mitosciences, USA). ..

other:

Article Title: MAP Kinase Phosphatase-5 Deficiency Improves Endurance Exercise Capacity.
Article Snippet: OXPHOS antibody cocktail (MS604) was obtained from MitoSciences (Waltham, MA, USA).

Article Title: Cannabidiol protects C2C12 myotubes against cisplatin-induced atrophy by regulating oxidative stress.
Article Snippet: Cancer and chemotherapy induce a severe loss of muscle mass (known as cachexia), which negatively impact cancer treatment and patient survival.. The aim of the present study was to investigate whether cannabidiol (CBD) administration may potentially antagonize the effects of cisplatin in inducing muscle atrophy, using a model of myotubes in culture.. Cisplatin treatment resulted in a reduction of myotube diameter (15.7 ± 0.3 vs. 22.2 ± 0.5 lm, P < 0.01) that was restored to control level with 5 lM CBD (20.1 ± 0.4 lM, P < 0.01).

Article Title: Defective brown adipose tissue thermogenesis and impaired glucose metabolism in mice lacking Letmd1
Article Snippet: Primary antibodies used in this study were anti-Letmd1 (LSBio; LS-C335200), anti-Brg1 (Bethyl; A300-813A-T or Cell Signaling; 49360S), anti-Ucp1 (R&D systems; MAB6158), anti-Prdm16 (R&D systems; AF6295), anti-Ebf2 (R&D systems; AF7006), anti-OXPHOS (MitoSciences; MS604), anti-lamin (Santa Cruz Biotechnology; sc-376248), anti-VDAC (Cell Signaling; 4661T), anti-FABP4 (Santa Cruz; sc-271529), anti-Flag (GenScript; A00187), anti-V5 (ThermoFisher; R96025), anti-β-actin (Santa Cruz; sc-47778), anti-Gapdh (Santa Cruz; sc-25778), and anti-vinculin (Sigma, V9131).

Article Title: Sulfur Amino Acid Restriction alters mitochondrial function depending on tissue, sex, and Methionine sulfoxide reductase A (MsrA) status.
Article Snippet: Antibody used: Total OxPhos/MitoPro le (MitoSciences, MS604/AbCam, ab110413).



Similar Products

90
MitoSciences total oxphos rodent antibody cocktail ms604
Total Oxphos Rodent Antibody Cocktail Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pm40616270-68-35-39
Average 90 stars, based on 1 article reviews
total oxphos rodent antibody cocktail ms604 - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

90
MitoSciences primary antibodies for oxidative phosphorylation complexes ms604
Primary Antibodies For Oxidative Phosphorylation Complexes Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pm40387167-77-33-36
Average 90 stars, based on 1 article reviews
primary antibodies for oxidative phosphorylation complexes ms604 - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

90
MitoSciences oxphos antibody cocktail ms604
Oxphos Antibody Cocktail Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pm40136658-59-0-7
Average 90 stars, based on 1 article reviews
oxphos antibody cocktail ms604 - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

90
MitoSciences oxphos cocktail ms604
Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an <t>OXPHOS</t> <t>antibody</t> <t>cocktail.</t> Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.
Oxphos Cocktail Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pmc11795799-78-69-72
Average 90 stars, based on 1 article reviews
oxphos cocktail ms604 - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

90
MitoSciences total oxphos antibody ms604
Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an <t>OXPHOS</t> <t>antibody</t> <t>cocktail.</t> Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.
Total Oxphos Antibody Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pm38406827-69-1-9
Average 90 stars, based on 1 article reviews
total oxphos antibody ms604 - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

90
MitoSciences total oxphos/mitopro le ms604
Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an <t>OXPHOS</t> <t>antibody</t> <t>cocktail.</t> Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.
Total Oxphos/Mitopro Le Ms604, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/ppr0777326-83-3-5
Average 90 stars, based on 1 article reviews
total oxphos/mitopro le ms604 - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

86
Cell Signaling Technology Inc ms604
Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an <t>OXPHOS</t> <t>antibody</t> <t>cocktail.</t> Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.
Ms604, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/pm37196697-432-12-19
Average 86 stars, based on 1 article reviews
ms604 - by Bioz Stars, 2026-09
86/100 stars
  Buy from Supplier

90
MitoSciences denaturing form of oxphos complexes cocktail ms604 antibody
Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an <t>OXPHOS</t> <t>antibody</t> <t>cocktail.</t> Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.
Denaturing Form Of Oxphos Complexes Cocktail Ms604 Antibody, supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/bio_rxiv__2023__03__30__534893-282-49-54
Average 90 stars, based on 1 article reviews
denaturing form of oxphos complexes cocktail ms604 antibody - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

90
MitoSciences oxphos (mitoprofile total oxphos protein, #ms604)
Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an <t>OXPHOS</t> <t>antibody</t> <t>cocktail.</t> Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.
Oxphos (Mitoprofile Total Oxphos Protein, #Ms604), supplied by MitoSciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ms604/mitoprofile+total+oxphos+rodent+wb+antibody+cocktail/pmc03986237-96-115-114
Average 90 stars, based on 1 article reviews
oxphos (mitoprofile total oxphos protein, #ms604) - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

Image Search Results


Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an OXPHOS antibody cocktail. Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.

Journal: Heliyon

Article Title: USP14 is crucial for proteostasis regulation and α-synuclein degradation in human SH-SY5Y dopaminergic cells

doi: 10.1016/j.heliyon.2025.e42031

Figure Lengend Snippet: Oxidative stress with elongation of mitochondria in USP14 deficient cells. Control and USP14-deleted cells were cultured and analyzed as indicated below. (A) Live-cell imaging using MitoTracker DeepRed FM and a 100× objective of Nikon-Eclipse Ti-E inverted wide-field microscope equipped with an environmental chamber. MitoTracker DeepRed FM channel (644/665 nm) represented in grayscale. Bottom panels, higher magnification. Left-bottom panel, quantification of mitochondrial branch length in μM from 90 to 100 control and USP14 deleted cells. Scale bar: 100 μM. Typical experiment is shown and was repeated three times with similar results. (B) EM imaging was done as described in Methods. Note elongated mitochondria (MT) in USP14 deleted cells. Red stars ∗ mark the presence of electron-dense lysosomes/autophagosomes/vesicles that were increased in the USP14-deleted cells compared with controls. N represents nuclear compartment. The experiment was repeated with similar results. (C) Left panel, immunoblot using an OXPHOS antibody cocktail. Right panel, quantification of the densitometry ratio of CI-CV subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1 and CV-ATP5A) normalized to β-actin. Values are means ± S.E.M. ∗p ≤ 0.05, ns = not significant. n = 3. p-value was calculated by Student's t-test. (D) Live-cell imaging using CellROX Green and a 20× objective of EVOS FL microscope. Control cells treated with 5 μM H 2 O 2 for 90 min served as a positive control for oxidative stress. Top, CellROX Green. Bottom, phase-contrast images. Scale bar: 200 μM. Typical experiment is shown and was repeated three times with similar results.

Article Snippet: The primary antibodies used included: USP14 (J6111-6D6, Sigma, Germany), α-synuclein (2642, CST, USA), pS129-α-synuclein (EP1536Y, Abcam, GB), 20S CP subunits cocktail (BML-PW8195, Enzo Lifesciences, USA), K48-linkage specific polyubiquitin (4289, CST), PSMD2 (sc-271775, SantaCruz, USA), PSMC2 (14395, CST), UCHL5 (sc-271002, SantaCruz, USA), TFEB (37785, CST, USA), GBA1 (sc-166407, SantaCruz, USA), GABARAP (13733, CST, USA), LC3B (3868, CST, USA), p62/SQSTM1 (P0067, Sigma, Germany), beta-Actin (A2066, Sigma, Germany), GAPDH (MAB374, Millipore, Germany), OXPHOS cocktail (MS604, Mitosciences, USA – Currently Abcam, GB).

Techniques: Control, Cell Culture, Live Cell Imaging, Microscopy, Imaging, Western Blot, Positive Control