Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Pressure overload-induced cardiac remodeling and dysfunction in the absence of interleukin 6 in mice.

doi: 10.1038/labinvest.2012.97

Figure Lengend Snippet: Figure 3 IL-6 deletion was insufficient to suppress apoptosis and had no effect on autophagy in pressure-overloaded hearts. (a) There was no group difference in TUNEL-positive cardiac myocytes in CON and IL6KO mice 2 weeks after TAC (P ¼ 0.83). (b) Caspase 3/7 activity was not significantly different in LV samples from CON and IL6KO 2 weeks after TAC (P ¼ 0.90). (c) Representative western blots showing protein contents of Bcl2, cathepsin D and LC3-II in LV samples from CON and IL6KO mice before and 2 weeks after TAC. There was no difference in protein content of Bcl2 (P ¼ 0.55) (d), cathepsin D (P ¼ 0.31) (e) and LC3-II (P ¼ 0.64) (f) in LV samples from CON and IL6KO mice 2 weeks after pressure overload. Probability values are from CON vs IL6KO comparison by Bonferroni post hoc test (CON vs IL6KO, 2 weeks after TAC) after two-way ANOVA. Error bars denote 1 s.e.m.; numbers in bars indicate group size.

Article Snippet: LV homogenates were used for western blotting as previously described,20 using the following antibodies: Bcl2 (Cat #sc-7382, Santa Cruz Biotechnology, Santa Cruz, CA, USA), a-smooth muscle actin (Cat #A5228, Sigma, St Louis, MO, USA), periostin (Cat #AF2955, R&D Systems, Minneapolis, MN, USA), cathepsin D (Cat #AF1029, R&D Systems), LC3 (Cat #2775, Cell Signaling Technology, Danvers, MA, USA), phospho-STAT3 (Cat #9145, Cell Signaling Technology), and STAT3 (Cat #9139, Cell Signaling Technology).

Techniques: TUNEL Assay, Activity Assay, Western Blot, Comparison

Journal: Molecular Medicine

Article Title: Melatonin ameliorates age-related sarcopenia by inhibiting fibrogenic conversion of satellite cell

doi: 10.1186/s10020-024-00998-2

Figure Lengend Snippet: Melatonin reduced muscle fibrosis in old mice. A , B Representative images of sirius red staining of gastrocnemius muscle (Scale bar: 200 µm). C , D Representative images of immunofluorescence staining showing COL-I (green) + DAPI (blue) of gastrocnemius muscle slides (Scale bar: 100 µm). E Representative western blotting. F – J Densitometric analysis of FN, COL-I, COL-III, Vim, and a-SMA in gastrocnemius muscle. The densities were normalized with those of β-Actin. Data represent mean ± S.D. of at least three independent experiments (n = 6 per group). *p < 0.05 versus Young. # p < 0.05 versus Old

Article Snippet: Then, the membrane was incubated overnight at 4 °C with primary antibodies (myogenin (MyoG) (1:100, DSHB Cat# AB_2146602, USA), myoblast determination protein 1 (MyoD) (1:100, DSHB Cat# D7F2, USA), Myosin Heavy Chain (MyHC) (1:100, DSHB Cat# MF 20, USA), Paired Box 7 (Pax7)(1:100, DSHB Cat# pax7, USA), P53(1:1000, Abcam Cat# ab131442, UK), p16INK4a(1:1000, Abcam Cat# ab189034, UK), Collagen I (1:1000, Abcam Cat# ab270993, UK), Collagen III(1:1000, Abcam Cat# ab184993, UK), α-smooth muscle actin(α-SMA) (1:1000, Abmart Cat# T51132S, China), Fibronectin(1:1000, Abmart Cat# PA3222M, China), Vimentin (1:500, Santa Cruz Cat# sc-66002, USA), TGF-β1 (1:1000, Affinity Biosciences Cat# #AF1027, USA), Smad2/3 (1:1000, Affinity Biosciences Cat# AF6367, USA) and p-Smad2/3 (Thr8) (1:1000, Affinity Biosciences Cat# AF3367, USA)).

Techniques: Staining, Immunofluorescence, Western Blot

Journal: Molecular Medicine

Article Title: Melatonin ameliorates age-related sarcopenia by inhibiting fibrogenic conversion of satellite cell

doi: 10.1186/s10020-024-00998-2

Figure Lengend Snippet: Melatonin inhibited fibrogenic differentiation in aging SCs. The SCs were categorized into three groups and culture in differentiate medium: Control group, Group treated with 30 mg/ml d -gal, and Group treated with both 30 mg/ml d -gal and 100 µM melatonin. A Representative image of immunofluorescence staining of FN (red) and nucleus DAPI (blue) in each group of cells after 24 h differentiation (Scale bar: 50 µm). B Analyses of the proportion of FN positive nuclei. C – F Representative western blotting and densitometric analysis of FN, Vim, and a-SMA in each group of cells after 24 h differentiation. G Representative image of immunofluorescence staining of COL-I (orange) and nucleus DAPI (blue) in each group of cells after 72 h differentiation (Scale bar: 20 µm). H Analysis of the COL-I positive area ratio in differentiated SCs. I – K Representative western blotting and densitometric analysis of COL-I and COL-III in each group of cells after 72 h differentiation. Data represent mean ± S.D. of at least three independent experiments (n = 3 per/group). *p < 0.05, **p < 0.01 versus CON, # p < 0.05, ## p < 0.01 versus d -gal

Article Snippet: Then, the membrane was incubated overnight at 4 °C with primary antibodies (myogenin (MyoG) (1:100, DSHB Cat# AB_2146602, USA), myoblast determination protein 1 (MyoD) (1:100, DSHB Cat# D7F2, USA), Myosin Heavy Chain (MyHC) (1:100, DSHB Cat# MF 20, USA), Paired Box 7 (Pax7)(1:100, DSHB Cat# pax7, USA), P53(1:1000, Abcam Cat# ab131442, UK), p16INK4a(1:1000, Abcam Cat# ab189034, UK), Collagen I (1:1000, Abcam Cat# ab270993, UK), Collagen III(1:1000, Abcam Cat# ab184993, UK), α-smooth muscle actin(α-SMA) (1:1000, Abmart Cat# T51132S, China), Fibronectin(1:1000, Abmart Cat# PA3222M, China), Vimentin (1:500, Santa Cruz Cat# sc-66002, USA), TGF-β1 (1:1000, Affinity Biosciences Cat# #AF1027, USA), Smad2/3 (1:1000, Affinity Biosciences Cat# AF6367, USA) and p-Smad2/3 (Thr8) (1:1000, Affinity Biosciences Cat# AF3367, USA)).

Techniques: Control, Immunofluorescence, Staining, Western Blot

Journal: Molecular Medicine

Article Title: Melatonin ameliorates age-related sarcopenia by inhibiting fibrogenic conversion of satellite cell

doi: 10.1186/s10020-024-00998-2

Figure Lengend Snippet: Melatonin suppressed TGF-β1/Smad2/3 Signaling in old Mice and d -gal induced aging SCs. A Representative image of immunofluorescence staining of TGF-β1/Smad2/3 (red) and nucleus DAPI (blue) of gastrocnemius muscle slides (Scale bar: 100 µm). B – D Representative western blotting and densitometric analysis of TGF-β1, Smad2/3, and p-Smad2/3 in gastrocnemius muscle of young mice, old mice, and melatonin-treated old mice. E – G SCs were differentiate 24 h in Control group, Group treated with 30 mg/ml d -gal, and Group treated with both 30 mg/ml d -gal and 100 µM melatonin. Representative western blotting and densitometric analysis of FN, Vim, and a-SMA in each group of cells. Data represent mean ± S.D. of at least three independent experiments. C , D *p < 0.05 versus Young. # p < 0.05, versus Old, (n = 6 per/group). F , G *p < 0.05 versus CON, # p < 0.05 versus d -gal, (n = 3 per/group)

Article Snippet: Then, the membrane was incubated overnight at 4 °C with primary antibodies (myogenin (MyoG) (1:100, DSHB Cat# AB_2146602, USA), myoblast determination protein 1 (MyoD) (1:100, DSHB Cat# D7F2, USA), Myosin Heavy Chain (MyHC) (1:100, DSHB Cat# MF 20, USA), Paired Box 7 (Pax7)(1:100, DSHB Cat# pax7, USA), P53(1:1000, Abcam Cat# ab131442, UK), p16INK4a(1:1000, Abcam Cat# ab189034, UK), Collagen I (1:1000, Abcam Cat# ab270993, UK), Collagen III(1:1000, Abcam Cat# ab184993, UK), α-smooth muscle actin(α-SMA) (1:1000, Abmart Cat# T51132S, China), Fibronectin(1:1000, Abmart Cat# PA3222M, China), Vimentin (1:500, Santa Cruz Cat# sc-66002, USA), TGF-β1 (1:1000, Affinity Biosciences Cat# #AF1027, USA), Smad2/3 (1:1000, Affinity Biosciences Cat# AF6367, USA) and p-Smad2/3 (Thr8) (1:1000, Affinity Biosciences Cat# AF3367, USA)).

Techniques: Immunofluorescence, Staining, Western Blot, Control