α ubc9  (Proteintech)

Journal: BMC Cancer

Article Title: FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling

doi: 10.1186/s12885-025-15364-6

Figure Lengend Snippet: FOXL2 SUMOylation intricately regulates increased expression in CAFs. ( A ) SUMOylation sites of FOXL2 was analyzed by SUMOplot™ analysis. Shown are the top 7 predicted lysine residues; ( B ) In vitro sumoylation assay was employed to confirm the predicted SUMOylation sites using HA-tagged wild-type (WT), or the K25R, K87R, K114R, K150R, and 4KR (where K25, K87, K114 and K150 were all mutated to R). Shown is a representative blot and densitometry analysis of SUMOylated-FOXL2/Total FOXL2; ( C ) The association between SUMOylation and FOXL2 stability was determined by western blotting in HEK-293T cells using α-HA. Cells were transfected with either HA-FOXL2-WT or HA-FOXL2-K25/87R (double mutant, 2KR). The membrane was stripped and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( D ) Cells were transfected with HA-FOXL2-WT or, HA-FOXL2-2KR ± His-SUMO1. CHX (100 µg/ml) was added to inhibit translation allowing tracking of FOXL2 stability in the presence and absence of His-SUMO1. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot and densitometry analysis of three technical replicates; ( E ) DMSO (control), MG132 (proteasome inhibitor) or chloroquine (lysosome inhibitor) were added into HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, before CHX (100 µg/ml) was added. Blots were probed with α-HA and re-probed with GAPDH to confirm equivalent loading. Shown is a representative blot; ( F ) IP assay was used to test the association between FOXL2 SUMOylation and ubiquitination. Lysates obtained from HEK-293T cells transfected with HA-FOXL2-WT or HA-FOXL2-2KR, along with FLAG-UBC9, were immunoprecipitated using α-HA antibody and then probed with α-FLAG antibody. Shown is a representative blot; *, *** P < 0.05, P < 0.001

Article Snippet: The different primary antibodies used were α-SUMO1 (67559-1-Ig, 1:3000), α-UBC9 (10070-1-AP, 1:2000), α-Vimentin (10366-1-AP, 1:5000), α-N-cadherin (22018-1-AP, 1:5000), α-E-cadherin (20874-1-AP, 1:20000) (Proteintech, Wuhan, China); α-HA (A02041, 1:800), α-His (A02050, 1:800), α-Flag (A02010, 1:800) (Abbkine, Wuhan, China); α-FOXL2 (ab246511, 1:1000), α-Snail (ab216347, 1:1000) (Abcam, Cambridge, MA, USA); and, α-Smad2/3 (D7G7, 1:1000), α-p-Smad2 (Ser465/467)/Smad3 (Ser423/425) (D27F4, 1:1000) (CST, Boston, MA, USA).

Techniques: Expressing, In Vitro, Western Blot, Transfection, Mutagenesis, Membrane, Control, Ubiquitin Proteomics, Immunoprecipitation

Journal: BMC Cancer

Article Title: FOXL2 + cancer-associated fibroblasts enhances epithelial ovarian cancer development via TGFβ/Smad signaling

doi: 10.1186/s12885-025-15364-6

Figure Lengend Snippet: FOXL2 SUMOylation requires SUMO1 and UBC9/UBE2I. ( A ) Putative interaction of FOXL2 with SUMO1, SUMO2, SUMO3, and SUMO4 was detected by String analysis ( https://cn.string-db.org/ ); ( B ) In vitro sumoylation assay was employed to confirm String analysis’s prediction of SUMO1, HEK-293T cells were transfected with HA-FOXL2-WT, FLAG-UBC9, and His-SUMO1-4. In vitro SUMOylation assay using Ni 2+ -NTA pull-down determined that FOXL2 was mainly modified by SUMO1. Shown is a representative blot; ( C ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation. HEK-293T cells were transfected with HA-FOXL2-WT and His-SUMO1 ± FLAG-UBC9. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot; ( D ) In vitro sumoylation assay was employed to determine whether UBC9 is compulsorily required for FOXL2 SUMOylation in CAFs. CAFs were transduced using either a non-targeting control shRNA or shRNA targeting UBC9 . Transduced cells were transfected with HA-FOXL2-WT and His-SUMO1. In vitro SUMOylation using Ni 2+ -NTA pull-down assay determined that UBC9 is required for FOXL2 SUMOylation. Shown is a representative blot

Article Snippet: The different primary antibodies used were α-SUMO1 (67559-1-Ig, 1:3000), α-UBC9 (10070-1-AP, 1:2000), α-Vimentin (10366-1-AP, 1:5000), α-N-cadherin (22018-1-AP, 1:5000), α-E-cadherin (20874-1-AP, 1:20000) (Proteintech, Wuhan, China); α-HA (A02041, 1:800), α-His (A02050, 1:800), α-Flag (A02010, 1:800) (Abbkine, Wuhan, China); α-FOXL2 (ab246511, 1:1000), α-Snail (ab216347, 1:1000) (Abcam, Cambridge, MA, USA); and, α-Smad2/3 (D7G7, 1:1000), α-p-Smad2 (Ser465/467)/Smad3 (Ser423/425) (D27F4, 1:1000) (CST, Boston, MA, USA).

Techniques: In Vitro, Transfection, Modification, Pull Down Assay, Control, shRNA

Journal: Molecular Metabolism

Article Title: Ube2i deletion in adipocytes causes lipoatrophy in mice

doi: 10.1016/j.molmet.2021.101221

Figure Lengend Snippet: Generation of conditional Ube2i gene deletion mice. (A) Ube2i fl/fl mice were generated using CRISPR/Cas9 gene editing. Exons 3 and 4 of the Ube2i ( Ubc9 ) gene were targeted by sgRNAs designed complementary to intronic sequences flanking the exons, then loxP sequences were introduced by DNA donor oligonucleotides. LoxP sites were inserted before exon 3 and after exon 4 (black arrows). Primers detect the 5’ (P1, P2) and 3’ (P3, P4) loxP sequences. (B) PCR analyses of floxed alleles at the targeted loci in genomic DNA extracted from ear clips of wild-type (+/+), fl/+, and fl/fl mice. PCR products were run on agarose gels with expected band sizes for P1–P2: wild-type (+) 319 bp and loxP allele (fl) 353 bp and P3–P4: wild-type (+) 427 bp and loxP allele (fl) 461 bp. The image shows a wild-type control #1107 +/+, founder heterozygous mouse #974 fl/+, and homozygous F2 offspring #1091 fl/fl. (C) Western blot analysis of UBE2I expression in inguinal WAT (iWAT) derived fibroblasts from Ube2i fl/fl mice after transduction with adenoviral GFP or Cre. To determine cell autonomous effects of Ube2i deletion in adipocytes, tamoxifen inducible Cre-expressing mice ( CAG-Cre ) were crossed with Ube2i fl/fl mice. iWAT SVF cells were isolated from CAG-Cre;Ube2i fl/fl and Ube2i fl/fl control mice. All cells were treated with tamoxifen to induce Cre recombination followed by adipocyte differentiation (diff) for eight days. (D) Western blot analysis and (E) relative gene expression by qPCR were performed to assess Ube2i depletion, adipocyte maturation, and beige fat cell markers. Gray = Ube2i fl/fl , yellow = CAG-Cre;Ube2i fl/fl . Data are presented as mean +/− SEM, ∗p < 0.05. (F) Brightfield and fluorescent images of differentiated Ube2i fl/fl and CAG-Cre;Ube2i fl/fl cells stained for lipids (green, LipidTOX), perilipin (red), and nuclei (blue, DAPI). Scale bars 50 μm. (G) Whole cell lysates from differentiated cells were subjected to Western blot analysis of cleaved (p41/43, p18) and uncleaved Caspase-8. (H) Strategy for generating adipocyte-specific Ube2i gene deletion. Ube2i fl/fl mice were bred with Adipoq-Cre mice to generate adipocyte-specific Ube2i knockout ( Ube2i a-KO ) and Ube2i fl/fl (control) mice. (I) To validate gene deletion of Ube2i , genomic DNA was extracted from Ube2i a-KO and Ube2i fl/fl gonadal WAT (gWAT) samples and PCR products were run on an agarose gel to detect the 5’ (P1, P2 primers) and 3’ (P3, P4 primers) loxP sequences, as well as a product that spans exons 3–4 (P1+P2+P4; 1597 bp, green arrow) or the deletion product (509 bp, red arrow). (J) Western blots of whole tissue lysates from iWAT and BAT of seven-day-old mice were probed for UBE2I and adiponectin (ADIPOQ), and HSP90 served as the invariant loading control.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-UBE2I (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), α-PPARγ (Cell Signaling #2443), α-Caspase-8 (Cell Signaling #4790), and α-Cleaved Caspase-8 (Cell Signaling #8592).

Techniques: Generated, CRISPR, Control, Western Blot, Expressing, Derivative Assay, Transduction, Isolation, Gene Expression, Staining, Knock-Out, Agarose Gel Electrophoresis

Journal: Molecular Metabolism

Article Title: Ube2i deletion in adipocytes causes lipoatrophy in mice

doi: 10.1016/j.molmet.2021.101221

Figure Lengend Snippet: Adipocyte-specific Ube2i deletion impairs WAT expansion in male and female mice. ( A) Body weight and (B) WAT weights (g) in combined male and female mice at 7 (n = 5–9), 30 (n = 4/group), 60 (n = 4/group), and 180 (n = 23–24/group) days of age for Ube2i fl/fl (gray) and Ube2i a-KO (purple) mice. Data are presented as mean +/− SEM; ∗p < 0.05. (C) Relative gene expression by qPCR in gWAT (left, yellow) and iWAT (right, cyan) from Ube2i a-KO (purple) and Ube2i fl/fl control (gray) mice for adipocyte maturation, lipid metabolism, inflammatory, and senescence genes represented as a heatmap of z-scores. Gray heatmap squares indicate outliers excluded based on Grubbs' test. (D) Representative H/E stained gWAT and iWAT sections from one-month-old Ube2i a-KO and Ube2i fl/fl control mice. Scale bar 100 μm. (E) Ube2i a-KO and Ube2i fl/fl control mice were weighed for up to 23 weeks in males (n = 12–14/group, mean +/− SD) and females (n = 6–8/group, mean +/− SD). (F) Assessment of fat and lean mass (% body weight (left) and in grams (right); male n = 11–15/group, female n = 5–7/group). (G) Tissue weights (% body weight; male n = 15-16/group, female n = 8/group) at necropsy. (H) Corresponding necropsy images from six-month-old Ube2i fl/fl and Ube2i a-KO mice. Images of excised tissues demonstrate gross morphological increases in liver size, reductions in iWAT and gWAT, and lighter coloring of liver and BAT. (E – H) Gray = Ube2i fl/fl male and female controls, blue = male Ube2i a-KO , red = female Ube2i a-KO . Data are presented as mean +/− SEM; ∗p < 0.05.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-UBE2I (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), α-PPARγ (Cell Signaling #2443), α-Caspase-8 (Cell Signaling #4790), and α-Cleaved Caspase-8 (Cell Signaling #8592).

Techniques: Gene Expression, Control, Staining

Journal: Molecular Metabolism

Article Title: Ube2i deletion in adipocytes causes lipoatrophy in mice

doi: 10.1016/j.molmet.2021.101221

Figure Lengend Snippet: Ube2i a-KO mice display WAT dysfunction with increased inflammation and apoptosis. (A) H/E-stained sections from gWAT and iWAT of male (top row) and female (bottom row) mice show substantial immune and stromal cell infiltration in Ube2i a-KO mice. Scale bar 100 μm. (B) Fed serum adiponectin (ng/ml), leptin (ng/ml), and free fatty acids (μM) in male (blue/gray; n = 11–12/group) and female (red/gray; n = 7–9/group) Ube2i a-KO mice compared to Ube2i fl/fl controls. Data are presented as mean +/− SEM; ∗p < 0.05. (C) Relative gene expression by qPCR in gWAT (left) and iWAT (right) from male (blue) and female (red) Ube2i a-KO (pink) and Ube2i fl/fl control (green) mice for adipocyte maturation, lipid metabolism, inflammatory, and senescence genes represented as a heatmap of z-scores (∗p < 0.05, #p < 0.10). Gray heatmap squares indicate outliers excluded based on Grubbs' test. (D) Tissue lysates from gWAT (left) and iWAT (right) of Ube2i fl/fl and Ube2i a-KO mice were subjected to Western blot analysis of cleaved (p41/43, p18) and uncleaved Caspase-8. All analyses were performed in six-month-old adult mice.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-UBE2I (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), α-PPARγ (Cell Signaling #2443), α-Caspase-8 (Cell Signaling #4790), and α-Cleaved Caspase-8 (Cell Signaling #8592).

Techniques: Staining, Gene Expression, Control, Western Blot

Journal: Molecular Metabolism

Article Title: Ube2i deletion in adipocytes causes lipoatrophy in mice

doi: 10.1016/j.molmet.2021.101221

Figure Lengend Snippet: Adipocyte-specific Ube2i knockout mice develop insulin resistance and hepatic steatosis. ( A) Representative H/E stained liver sections from male (top row) and female (bottom row) Ube2i a-KO and Ube2i fl/fl control mice show lipid droplet accumulation in Ube2i a-KO mice. Scale bar 100 μm. (B) Hepatic triglycerides (TGs) per gram liver tissue (n = 7/group). (C) Fed serum FGF21 (pg/ml) (male n = 11–12/group; female n = 7–9/group). (D) Fed serum glucose (mg/dl) and (E) insulin (ng/ml) (male n = 11–12/group; female n = 7–9/group). (F) Insulin and (G) glucose tolerances tests were performed on Ube2i fl/fl and Ube2i a-KO male (n = 11–15/group) and female (n = 5–7/group) mice. Area under the curve is also shown. Values labeled on panels in (F) indicate the percentage decrease from initial fasting blood glucose. (H) Serum insulin during glucose tolerance test from Ube2i fl/fl and Ube2i a-KO male (n = 5/group) and female (n = 4–6/group) mice. Gray = Ube2i fl/fl male and female controls, blue = male Ube2i a-KO , red = female Ube2i a-KO . Data are presented as mean +/− SEM; ∗p < 0.05 between groups, #p < 0.05 versus time zero. All analyses were performed in six-month-old adult mice.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-UBE2I (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), α-PPARγ (Cell Signaling #2443), α-Caspase-8 (Cell Signaling #4790), and α-Cleaved Caspase-8 (Cell Signaling #8592).

Techniques: Knock-Out, Staining, Control, Labeling

Journal: Molecular Metabolism

Article Title: Ube2i deletion in adipocytes causes lipoatrophy in mice

doi: 10.1016/j.molmet.2021.101221

Figure Lengend Snippet: Adipocyte-specific Ube2i deletion increases energy expenditure and cold intolerance. (A) H/E stained BAT sections from male and female Ube2i fl/fl and Ube2i a-KO mice show large unilocular lipid droplets in Ube2i a-KO mice. Scale bar 100 μm. (B) Relative gene expression by qPCR in BAT from male (left, blue) and female (right, red) Ube2i a-KO (pink) and Ube2i fl/fl control (green) mice for brown adipocyte and lipid metabolism genes represented as a heatmap of z-scores (∗p < 0.05, #p < 0.10). Six-month-old Ube2i fl/fl (gray) and Ube2i a-KO (blue) male mice were individually housed and monitored in CLAMS home cages for 6 days (n = 4–5/group). (C) Recorded traces of energy expenditure (kcal/hour) with mean values for dark/light periods (kcal/h). (D) Average RER during dark and light periods. (E) Activity was measured by wheel running. (F) Recorded traces of cumulative food intake (kcal) with mean values for dark/light periods (kcal) and (G) total food intake (kcal) per day. (H) Temperature probes were inserted under the skin to monitor intrascapular BAT temperature before (room temperature, RT) and after 2.5 h of cold (4 °C) exposure (n = 3–5/group). Statistical analysis of energy expenditure was performed by ANCOVA with lean body mass as a co-variate and cumulative food intake by standard ANOVA. Data are presented as mean +/− SEM; ∗p < 0.05, #p < 0.10.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-UBE2I (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), α-PPARγ (Cell Signaling #2443), α-Caspase-8 (Cell Signaling #4790), and α-Cleaved Caspase-8 (Cell Signaling #8592).

Techniques: Staining, Gene Expression, Control, Activity Assay

Journal: bioRxiv

Article Title: Ubc9 deletion in adipocytes causes lipoatrophy in mice

doi: 10.1101/2020.09.12.294629

Figure Lengend Snippet: (A) Ubc9 fl/fl mice were generated using CRISPR/Cas9 gene editing. Exons 3 and 4 of the Ube2i ( Ubc9 ) gene were targeted by sgRNAs designed complementary to intronic sequences flanking the exons, then loxP sequences were introduced by DNA donor oligonucleotides. LoxP sites were inserted before exon 3 and after exon 4 (black arrows). Primers detect the 5’ (P1, P2) and 3’ (P3, P4) loxP sequences. (B) PCR analyses of floxed alleles at the targeted loci in genomic DNA extracted from ear clips of wild-type (+/+), fl/+, and fl/fl mice. PCR products were run on agarose gels with expected band sizes for P1-P2: wild-type (+) 319 bp and loxP allele (fl) 353 bp and P3-P4: wild-type (+) 427 bp and loxP allele (fl) 461 bp. The image shows a wild-type control #1107 +/+, founder heterozygous mouse #974 fl/+, and homozygous F2 offspring #1091 fl/fl. (C) Western blot analysis of Ubc9 expression in iWAT SVF cells from Ubc9 fl/fl mice after transduction with adenoviral GFP or Cre. (D) Strategy for generating adipocyte-specific Ubc9 gene deletion. Ubc9 fl/fl mice were bred with AdipoQ-Cre mice to generate adipocyte-specific Ubc9 knockout ( Ubc9 a-KO ) and Ubc9 fl/fl (control) mice. (E) To validate deletion of Ubc9 , genomic DNA was extracted from Ubc9 a-KO and Ubc9 fl/fl gWAT samples and PCR products were run on an agarose gel to detect the 5’ (P1, P2 primers) and 3’ (P3, P4 primers) loxP sequences, as well as a product that spans exons 3-4 (P1+P2+P4; 1597 bp, green arrow) or the deletion product (509 bp, red arrow). (F) Western blots of whole tissue lysates from iWAT and BAT of seven-day old mice were probed for the indicated proteins. (G) Ubc9 a-KO and Ubc9 fl/fl (control) mice were weighed for up to 23 weeks in male (blue/gray; n=12-14, mean +/− SD) and female (red/gray; n=6-8, mean +/− SD).

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-Ubc9 (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), and α-Cre (Cell Signaling #7803).

Techniques: Generated, CRISPR, Control, Western Blot, Expressing, Transduction, Knock-Out, Agarose Gel Electrophoresis

Journal: bioRxiv

Article Title: Ubc9 deletion in adipocytes causes lipoatrophy in mice

doi: 10.1101/2020.09.12.294629

Figure Lengend Snippet: (A) Representative H/E stained liver sections from male (top row) and female (bottom row) Ubc9 a-KO and Ubc9 fl/fl control mice show lipid droplet accumulation in Ubc9 a-KO mice. Scale bar 100 μm. (B) Hepatic triglycerides (TGs) per gram liver tissue (n=7/group). (C) Fed serum FGF21 (pg/ml) (male, blue n=11-12; female, red n=7-9). (D) Fed serum glucose (mg/dl), (E) insulin (ng/ml), and (F) HOMA-IR measurements (male, blue n=11-12; female, red n=7-9). (G) Insulin and (H) glucose tolerances tests were performed on Ubc9 fl/fl and Ubc9 a-KO male (blue; n=11-15) and female (red; n=5-7) mice. Data represent mean +/− SEM; *p<0.05. Tissue and serum chemistry analyses were performed in six-month old adult mice.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-Ubc9 (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), and α-Cre (Cell Signaling #7803).

Techniques: Staining, Control

Journal: bioRxiv

Article Title: Ubc9 deletion in adipocytes causes lipoatrophy in mice

doi: 10.1101/2020.09.12.294629

Figure Lengend Snippet: Six-month old Ubc9 fl/fl (gray) and Ubc9 a-KO (blue) male mice were individually housed and monitored in CLAMS home cages for 6 days (n=4-5). Recorded traces of (A) energy expenditure (kcal/hour), (B) averaged RER during dark and light periods, and (C) cumulative food intake. Statistical analysis of energy balance was performed by ANCOVA with lean body mass as a co-variate. (D) Activity was measured by wheel running. (E) H/E stained BAT sections from male Ubc9 fl/fl and Ubc9 a-KO mice show large unilocular lipid droplets in Ubc9 a-KO mice. Scale bar 100 μm. (F) Relative gene expression by qPCR for markers of BAT function (n=4/group). (G) Temperature probes were inserted under the skin to monitor intrascapular BAT temperature before (room temperature, RT) and after 2.5 hours of cold (4°C) exposure (n=3-5). Data represent mean +/− SEM; *p<0.05, #p<0.10.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-Ubc9 (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), and α-Cre (Cell Signaling #7803).

Techniques: Activity Assay, Staining, Gene Expression

Journal: bioRxiv

Article Title: Ubc9 deletion in adipocytes causes lipoatrophy in mice

doi: 10.1101/2020.09.12.294629

Figure Lengend Snippet: (A) H/E stained sections from gonadal and inguinal WAT from male (top row) and female (bottom row) mice show substantial stromal cell infiltration in WAT of Ubc9 a-KO mice. Scale bar 100 μm. (B) Fed serum adiponectin (ng/ml), leptin (ng/ml), and free fatty acids (μM; FFA) in male (n=11-12) and female (n=7-9) Ubc9 a-KO mice compared to Ubc9 fl/fl controls. Relative gene expression by qPCR (n=4/group) for markers of mature adipocytes and inflammation in (C) gonadal and (D) inguinal WAT of male mice. Data represent mean +/− SEM; *p<0.05. All analyses were performed in six-month old adult mice.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-Ubc9 (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), and α-Cre (Cell Signaling #7803).

Techniques: Staining, Gene Expression

Journal: bioRxiv

Article Title: Ubc9 deletion in adipocytes causes lipoatrophy in mice

doi: 10.1101/2020.09.12.294629

Figure Lengend Snippet: (A) Assessment of fat and lean mass (% body weight; male n=11-15, female n=5-7) with (B) corresponding tissues weights (% body weight; male n=15-16, female n=8) and (C) necropsy images from six-month old Ubc9 fl/fl and Ubc9 a-KO mice. Images of excised tissues demonstrate gross morphological increases in liver size, reductions in iWAT and gWAT, and lighter coloring of the liver and BAT. (D) Body weight and (E) WAT weights (g) in male and female mice at 7 (n=5-9), 30 (n=4/group), 60 (n=4/group), and 180 (n=23-24/group) days of age. Data represent mean +/− SEM; *p<0.05.

Article Snippet: The following antibodies were used for immunoblotting: α-HSP90 (Cell Signaling #4877), α-Ubc9 (Cell Signaling #4786), α-ADIPOQ (Genetex #GTX112777), and α-Cre (Cell Signaling #7803).

Techniques: