Journal: Antioxidants

Article Title: Antioxidant Role and Cardiolipin Remodeling by Redox-Activated Mitochondrial Ca 2+ -Independent Phospholipase A 2 γ in the Brain

doi: 10.3390/antiox11020198

Figure Lengend Snippet: Antioxidant role of iPLA 2 γ in the cytosol and in vivo. Differences in protein carbonyl levels in brain homogenates ( A ) and interleukin-6 levels in serum ( B ) from WT mice and iPLA 2 γ-KO mice. Where indicated, mice were injected with LPS (5 mg/kg body weight) or iPLA 2 γ inhibitor r -BEL (1 mg/kg body weight) and LPS in parallel. * p < 0.05; ** p < 0.01; *** p < 0.005. ns, No significant differences were found between the rates in mitochondria isolated from iPLA 2 γ-KO mice.

Article Snippet: IL-6 levels were determined from collected sera using a commercially available kit (IL-6 Mouse ProQuantum Immunoassay Kit, Invitrogen, Waltham, MA, USA).

Techniques: In Vivo, Injection, Isolation

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: CD2F1 male mice were inoculated with 1 × 10 6 cxC26 or 0.5 × 10 6 ncxC26 cells. a Body weight, b tumor mass, c fat mass, and d lean mass post-implantation for mice injected with saline, cxC26 cells, or ncxC26 cells. n = 6 for saline, n = 8 for ncxC26, n = 12 for cxC26. The fat and lean mass were measured using Echo-MRI. e Mass of brown adipose tissue (BAT), gonadal white adipose tissue (gWAT), quadriceps (QD), heart, kidney, spleen, and liver on day 12 post implantation. n = 6 for saline, n = 9 for ncxC26, n = 5 for cxC26. f Muscle function of mice as evaluated using a grip strength meter. n = 6 for saline, n = 7 for ncxC26, n = 8 for cxC26. g Circulating cytokine profiles in saline, cxC26, and ncxC26-bearing mice. Mice were euthanized every two days from day 4 to day 12 for blood collection. The cytokine profiles were measured using Luminex 44 cytokine panel. n = 5∼6 biologically independent animals per group and time point. Cytokines highlighted in red are those with significantly altered levels in cxC26 compared to ncxC26 and saline (adjusted p-value < 0.01 by two-way ANOVA). h Circulating IL-6 concentration measured using a Proquantum immunoassay. n = 7 per group. All data in the figure are shown as the mean ± s.d. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA ( e, f ) or two-way ANOVA ( b, h ).

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Injection, Luminex, Concentration Assay

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: a Concentration of IL-6 in conditioned media of cxC26 scr (CRISPR/Cas9 scrambled gRNA control) and cxC26 IL-6 KOp with or without treatment of lipopolysaccharide (LPS) for 24 hours. b Body weight and c tumor volume post-tumor implantation. CD2F1 male mice were inoculated with 1 × 10 6 cxC26 or cxC26 IL-6 KOp cells. n = 4 per group. d Growth curve of cxC26, cxC26 scr and cxC26 IL-6 KOp cells in in vitro. n = 3 per group. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Concentration Assay, CRISPR, Tumor Implantation, In Vitro

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: a IL-6 level in conditioned media of cxC26 scr (CRISPR/Cas9 scrambled gRNA control) and cxC26 IL-6 KO subclones 1 (s1) and 2 (s2) with or without treatment of lipopolysaccharide (LPS) for 24 hours. (n = 4 per group) b-f CD2F1 male mice were inoculated with 1 × 10 6 cxC26 scr or 1 × 10 7 cxC26 IL-6 KO s1 cells. b Tumor growth and c body weight in saline, cxC26 scr or cxC26 IL-6 KO s1 tumor-bearing mice. d-f All groups were euthanized when the tumor-bearing mice developed cachexia (more than 15% of body weight loss). d Tumor weight, e muscle mass, and f circulating IL-6 concentration at terminal time point in saline, cxC26 scr and cxC26 IL-6 KO s1 tumor-bearing mice. b,c,d,f n = 4 for saline and for cxC26 IL-6 KO s1, n = 6 for cxC26. e n= 5 for saline and cxC26 IL-6 KO s1, n = 6 for cxC26. All data in the figure are shown as the mean ± s.d. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: CRISPR, Concentration Assay

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: CD2F1 male mice were inoculated with 1 × 10 6 cxC26 scr or 1 × 10 7 cxC26 IL-6 KO s2 cells. a Body weight, b terminal muscle mass, and c circulating IL-6 concentration at the end of experiment when the tumor-bearing animals developed cachexia (more than 15% of body weight loss). Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA. n = 5 for saline and cxC26 IL-6 KO s2, n = 6 for cxC26.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: Balb/c male mice were inoculated with 1 × 10 6 cxC26 scr or 1 × 10 7 cxC26 IL-6 KO s1 cells. a Body weight, b terminal muscle mass and c circulating IL-6 concentration at the end of experiment when the tumor-bearing animals developed cachexia (more than 15% of body weight loss). Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: CD2F1 Mice were inoculated 1 × 10 6 C26nci or C26nci LIF KO cells. All groups were euthanized when they developed cachexia phenotype (>15% of body weight loss). a Body weight and b mass of quadriceps at terminal time point. c Tumor growth. a-c n = 5 for saline, n = 6 for C26nci and C26nci LIF KO d Concentration of circulating LIF. e Concentration of circulating IL-6. d, e n = 4 for saline, n = 7 for C26nci, n = 5 for C26nci LIF KO. All data in the figure are shown as the mean ± s.d. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: cxC26 IL-6 receptor α ( IL-6Rα ) knock-out pool (KOp) was constructed using CRISPR/cas9. CD2F1 male mice were inoculated with 1 × 10 6 cxC26 or cxC26 IL-6Rα KOp cell. a Body weight curve and b terminal tumor weight at day 12. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by Student’s t-test. n = 4 for saline, n = 5 for cxC26 scr, n = 6 for cxC26 IL-6Rα KOp.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Knock-Out, Construct, CRISPR

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: NOD-SCID male mice were inoculated with 1 × 10 6 ncxC26 or cxC26 scr or cxC26 IL-6 KO cells. All groups were euthanized simultaneously on day 22. a Growth of tumors and b terminal tumor weight in ncxC26, cxC26 scr or cxC26 IL-6 KOs1 tumor-bearing mice. c The carcass weight at day 22, normalized by initial body weight. d Body weight e mass of quadriceps, and f concentration of circulating IL-6 at terminal time point. All data in the figure are shown as the mean ± s.d. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between groups by one-way ANOVA. n = 4 per group

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: IL-6 is dispensable for causing cachexia in the colon carcinoma 26 model

doi: 10.1101/2023.05.02.539076

Figure Lengend Snippet: Mice were inoculated 1 × 10 6 cxC26 scr or 1 × 10 7 cxC26 IL-6 KO s1 cells. The tumor-bearing mice were euthanized at the early phase (day 8 for all groups) when they had similar tumor size and terminal phase (day 13 for cxC26 and day 30 for cxC26 IL-6 KO s1) when they developed cachexia. The tumors were dissociated into single-cell suspension and stained with surface markers. All data are presented as percent of total live cells (7-AAD negative cells) and quantitate the proportion of CD45+ cells, B cells (CD45+/CD19+/CD11b-), CD4+ T-cells (CD45+/CD3+/CD4+/CD8-), CD8+ T-cells (CD45+/CD3+/CD4-/CD8+), macrophages (CD45+/CD11b+/F4/80+/Ly6G-), and neutrophils (CD45+/CD11b+/F4/80-/Ly6G+) were enumerated using flow cytometry. PD-1 positive cells were evaluated in CD4+ and CD8+ T-cells population. Statistical significance (t-test) between groups: *0.05 < p, **0.01 < p, *** 0.001 < p. n.s., non-significance. All data in the figure are shown as the mean ± s.d. Significance of the differences: *P < 0.05, ** P < 0.01, *** P < 0.001 between the cxC26 scr and cxC26 IL-6 KO s1 group by Student’s t-test. n = 5 per group.

Article Snippet: And circulating IL-6 was also determined using high sensitive IL-6 Mouse ProQuantum qPCR Immunoassay kit (Invitrogen) with real-time PCR.

Techniques: Staining, Flow Cytometry