Journal: JCI insight

Article Title: Prostate tumor-derived GDF11 accelerates androgen deprivation therapy-induced sarcopenia.

doi: 10.1172/jci.insight.127018

Figure Lengend Snippet: Figure 3. Castration increased active catabol- ic TGF-β family myokine proteins in skeletal muscles of tumor-bearing mice. (A–F) Protein expression in GAS muscle. (A) Representative immunoblots of soluble active myostatin (MSTN) C-terminal dimer and eukaryotic elongation factor 2 (EF2) expression in muscle from sets of 4 mice, castrated for the indicated times or sham castrat- ed. Lanes of immunoblots marked “C” contain identical control sample for interblot compari- son. (B) Quantification of relative MSTN levels for castrated mice (red) or sham-castrated mice (blue), from 3 determinations for each muscle (see Supplemental Figure 5 for additional immunoblots and supplemental materials for full, uncut gels). ELISA-determined protein levels of soluble active activin AA dimer (C), activin BB dimer (D), activin AB dimer (E), and soluble GDF11 (F), in muscle from 4 mice at each time point, measured 3 times each. (G–L) Protein expression in TRI muscle. (G) Representative immunoblot of MSTN and EF2 expression, as in A. (H) Quantification of MSTN levels, as in B. ELISA-determined protein levels of soluble active activin AA dimer (I), activin BB dimer (J), activin AB dimer (K), and soluble GDF11 (L), from 4 mice at each time point, measured 3 times each. Columns are sham-castrated normalized means at each time; bars are SEM. Individual mouse levels are indicated by open circles. *P < 0.05, **P < 0.01, and ***P < 0.001 versus sham-castrated group determined using 1-way ANOVA and Bonferroni’s correction (B and H) or Dunnett’s test (C–F and I–L).

Article Snippet: ELISA kits used were human/mouse/rat activin A ELISA kit (Quantikine DAC00B, R&D Systems, Bio-Techne), mouse activin B ELISA kit (E15932m, Cusabio Biotech Co., Ltd.), mouse activin AB and mouse GDF11 ELISA kits (CEA158Mu and SEC113Mu, respectively, Cloud-Clone Corp.), and total myostatin ELISA kit (Quantikine DGDF80, R&D Systems, Bio-Techne).

Techniques: Muscles, Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: JCI insight

Article Title: Prostate tumor-derived GDF11 accelerates androgen deprivation therapy-induced sarcopenia.

doi: 10.1172/jci.insight.127018

Figure Lengend Snippet: Figure 4. TGF-β family myokine ligand blockade reversed castration-induced sarcopenia. (A) Grip strength after castration or sham-castration of mice treated with PBS, ActRIIB-Fc, or anti-GDF11 antibody. (B) Dissected skeletal muscle mass 10 weeks after castra- tion by dissection and weighing. (C) Lean body mass by quantitative NMR (qNMR), as percentage of sham-cas- trated mice. Mean indicated as lines or columns, SEM as bars; n = 3–5/group, indicated by open circles. *P < 0.05 and **P < 0.01 for ActRIIB-Fc treated castrated versus vehicle-treated castrated mice determined using 2-way ANOVA and Tukey’s honestly significant differences (HSD) test. See Supplemental Figure 7 for additional information.

Article Snippet: ELISA kits used were human/mouse/rat activin A ELISA kit (Quantikine DAC00B, R&D Systems, Bio-Techne), mouse activin B ELISA kit (E15932m, Cusabio Biotech Co., Ltd.), mouse activin AB and mouse GDF11 ELISA kits (CEA158Mu and SEC113Mu, respectively, Cloud-Clone Corp.), and total myostatin ELISA kit (Quantikine DGDF80, R&D Systems, Bio-Techne).

Techniques: Dissection

Journal: JCI insight

Article Title: Prostate tumor-derived GDF11 accelerates androgen deprivation therapy-induced sarcopenia.

doi: 10.1172/jci.insight.127018

Figure Lengend Snippet: Figure 5. Castration increased GDF11 and myostatin in tumor and serum. (A–C) GDF11 and myostatin protein expression in tumor. (A) ELISA-determined levels of GDF11 in prostate tumor tissue, from 4 mice at each time, mea- sured 3 times. (B) Representative immunoblots of soluble active myostatin C-terminal dimer (MSTN) and eukaryotic elongation factor 2 (EF2) expression in prostate tumor tissue from sets of 4 mice, castrated for the indicated times or sham castrated. Lanes of immunoblots marked “C” contain identical control sample for interblot comparison. (C) Quantification of MSTN levels in tumor tissue from castrated (red) or sham-castrated (blue) mice, from 3 determi- nations for each tumor (see Supplemental Figure 5 for additional immunoblots and supplemental materials for full, uncut gels). (D–F) GDF11 and myostatin protein expression in serum. (D) ELISA-determined levels of GDF11 in serum, from 4 mice at each time, measured 3 times. (E) Representative immunoblots of MSTN expression in equal quanti- ties of serum protein from the 4 mice in A. (F) Quantification of MSTN levels in serum, as in B. (G–I) Comparison of GDF11 and myostatin protein expression between muscle and tumor. (G) ELISA-determined levels of GDF11 in GAS muscle and prostate tumor from sets of 4 mice, sham castrated (blue) or 2 weeks after castration (red), measured 3 times. (H) Representative immunoblots of MSTN in GAS muscle and prostate tumor from sets of 4 mice, sham-cas- trated or 8 weeks after castration. (I) Quantification of relative levels of MSTN between GAS muscle and prostate tumor. Columns are normalized means; bars are SEM. n = 4/group, indicated by open circles. *P < 0.05, **P < 0.01, and ***P < 0.001 versus sham-castrated group (except G and I, GAS muscle vs. tumor tissue), determined using 1-way ANOVA and Dunnett’s test (A, D, and G) or 1-way ANOVA and Bonferroni’s correction (C, F, and I).

Article Snippet: ELISA kits used were human/mouse/rat activin A ELISA kit (Quantikine DAC00B, R&D Systems, Bio-Techne), mouse activin B ELISA kit (E15932m, Cusabio Biotech Co., Ltd.), mouse activin AB and mouse GDF11 ELISA kits (CEA158Mu and SEC113Mu, respectively, Cloud-Clone Corp.), and total myostatin ELISA kit (Quantikine DGDF80, R&D Systems, Bio-Techne).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Comparison

Journal: JCI insight

Article Title: Prostate tumor-derived GDF11 accelerates androgen deprivation therapy-induced sarcopenia.

doi: 10.1172/jci.insight.127018

Figure Lengend Snippet: Figure 6. Model of ADT-induced catabolic TGF-β family myokine endocrine signaling in PrCa tumor–bearing mice. Graphs of castration-induced changes in myokine concentrations (colored lines) in skeletal muscle and serum of mice without prostate cancer (left side) or in skeletal muscle, serum, and tumor of mice with prostate cancer (right side). Tumor lobes are represented by the green (left lobe) and red (right lobe) circles. In adult tumor-free mice myostatin and the activins increase after castration but prior to strength and muscle mass loss. In PrCa tumor–bearing mice, tumor secretes GDF11 (blue) and later myostatin (red) into serum. Endocrine-derived GDF11 and myostatin increase in muscle prior to strength loss (early myostatin also moves from muscle into serum). The tumor-derived catabolic TGF-β family myokines exacerbate castration-induced sarcopenia in tumor-bearing mice.

Article Snippet: ELISA kits used were human/mouse/rat activin A ELISA kit (Quantikine DAC00B, R&D Systems, Bio-Techne), mouse activin B ELISA kit (E15932m, Cusabio Biotech Co., Ltd.), mouse activin AB and mouse GDF11 ELISA kits (CEA158Mu and SEC113Mu, respectively, Cloud-Clone Corp.), and total myostatin ELISA kit (Quantikine DGDF80, R&D Systems, Bio-Techne).

Techniques: Derivative Assay

Journal: Oncotarget

Article Title: Splenocytes derived from young WT mice prevent AD progression in APPswe/PSENldE9 transgenic mice

doi:

Figure Lengend Snippet: A-D. After SCs treatment, the levels of IFN-γ, IL-2, IL-4, IL-10 in brain of APPswe/PSENldE9 transgenic mice were assayed by ELISA. The content of the cytokines per mg brain tissue was calculated for statistical analysis. One way ANOVA with LSD post-hoc test was used for statistical analysis ( n = 6). E. The serum GDF11 level of APPswe/PSENldE9 transgenic mice with SCs treatments was assayed by ELISA. One way ANOVA with LSD post-hoc test was used for statistical analysis ( n = 6). F. The GDF11 expression of spleen isolated from old APPswe/PSENldE9 transgenic mice (14–15 month-old) and young mice (young, 6–8 week-old) mice was compared. Spleen homogenates were used for assaying the GDF11 expression in spleen by ELISA (Student's t -test for statistical analysis). Data are presented as mean ± SEM, n = 5, * p < 0.05.

Article Snippet: The GDF11 levels of serums and spleen homogenates were assayed by ELISA with a GDF11 ELISA kit (Cloud–Clone Corp., USA) according to the Corp's manual.

Techniques: Transgenic Assay, Enzyme-linked Immunosorbent Assay, Expressing, Isolation