Journal: Biology

Article Title: Free Fatty Acid Species Differentially Modulate the Inflammatory Gene Response in Primary Human Skeletal Myoblasts

doi: 10.3390/biology10121318

Figure Lengend Snippet: Mitochondrial abundance and activity in human primary myoblasts were differentially affected by FFAs. SkMs were treated with FAs conjugated with BSA or BSA alone (control). After 48 h, SkMs were incubated with MitoTracker Red ( a ) or Orange ( b ). Fluorescence images at identical settings and exposure times from 3 sets of experiments were simultaneously imported into the analysis software, processed to 8-bit images and analyzed. The integrated fluorescence intensity of FFA-treated cells was compared to that of control cells, which was set to 1 (c, dashed gray line). The results are presented as the mean ± SD of the relative fluorescence intensity of MitoTracker Red (white bars) or Orange (black bars). Statistical significance was calculated using one-way ANOVA with Bonferroni’s multiple comparison test. p ≤ 0.05 (*), p ≤ 0.01 (**).

Article Snippet: Primary human skeletal muscle myoblasts (SkMs) were obtained from Lonza, Basel, Switzerland (CC-2561).

Techniques: Activity Assay, Control, Incubation, Fluorescence, Software, Comparison

Journal: Biology

Article Title: Free Fatty Acid Species Differentially Modulate the Inflammatory Gene Response in Primary Human Skeletal Myoblasts

doi: 10.3390/biology10121318

Figure Lengend Snippet: FFAs do not affect the desmin expression pattern in human primary myoblasts. ( a ) SkMs were treated with FAs conjugated with BSA or BSA alone (control) before DAPI and desmin staining. ( b ) Fluorescence images from 3 sets of experiments were analyzed, as described in the legend to . The number of DAPI-positive FFA-treated cells was compared to that of control cells, which was set to 1 (c, dashed gray line). The data are presented as the mean ± SD of cell number (dotted bars). Statistical significance was calculated using one-way ANOVA with Bonferroni’s multiple comparison test. p ≤ 0.05 (*). p ≤ 0.01 (**).

Article Snippet: Primary human skeletal muscle myoblasts (SkMs) were obtained from Lonza, Basel, Switzerland (CC-2561).

Techniques: Expressing, Control, Staining, Fluorescence, Comparison

Journal: Biology

Article Title: Free Fatty Acid Species Differentially Modulate the Inflammatory Gene Response in Primary Human Skeletal Myoblasts

doi: 10.3390/biology10121318

Figure Lengend Snippet: FFAs are differentially utilized in human primary myoblasts. ( a ) SkMs were treated with FAs conjugated with BSA or BSA alone (control) for 48 h and stained using BODIPY. ( b ) Fluorescence images from 3 sets of experiments were analyzed as described in the legend to . The fluorescence intensity of FFA-treated cells was compared to that of control cells, which was set to 1 (c, dashed gray line). The results are presented as the mean ± SD of relative lipid accumulation (hatched gray bars). Statistical significance was calculated using one-way ANOVA with Bonferroni’s multiple comparison test. p ≤ 0.05 (*).

Article Snippet: Primary human skeletal muscle myoblasts (SkMs) were obtained from Lonza, Basel, Switzerland (CC-2561).

Techniques: Control, Staining, Fluorescence, Comparison

Journal: Biology

Article Title: Free Fatty Acid Species Differentially Modulate the Inflammatory Gene Response in Primary Human Skeletal Myoblasts

doi: 10.3390/biology10121318

Figure Lengend Snippet: FFAs selectively activate RTK phosphorylation in human primary myoblasts. SkMs were treated with FAs conjugated with BSA or BSA alone for two hours. Whole-cell lysates were prepared and each incubated with a single RTK array. Following incubation steps, the slides were washed, imaged and analyzed using a laser scanner. Fluorescence images were processed and analyzed simultaneously using analysis software. To obtain the relative phosphorylation (black bars) of individual RTKs indicated at the top of each diagram, integrated signal intensities from FFA-treated cells were compared to that from control cells, which was set to 1 ( a – g , dashed gray line). Data are presented as the mean ± SD of the relative phosphorylation. Statistical significance was calculated using one-way ANOVA with Bonferroni’s multiple comparison test. p ≤ 0.05 (*). p ≤ 0.01 (**).

Article Snippet: Primary human skeletal muscle myoblasts (SkMs) were obtained from Lonza, Basel, Switzerland (CC-2561).

Techniques: Phospho-proteomics, Incubation, Fluorescence, Software, Control, Comparison

Journal: The EMBO Journal

Article Title: TWEAK, via its receptor Fn14, is a novel regulator of mesenchymal progenitor cells and skeletal muscle regeneration

doi: 10.1038/sj.emboj.7601441

Figure Lengend Snippet: Human mesenchymal progenitor cells are a novel target cell type for TWEAK. (A) Human primary mesenchymal stem cells, skeletal muscle myoblasts, preadipocytes, chondrocytes and osteoblast precursors (Cambrex) were cultured according to the manufacturer's protocols. First-passage cells showed staining for TWEAK binding using Fc-TWEAK and for expression of Fn14 using the anti-hFn14 mAb ITEM-4. Anti-mouse and anti-human Fcs were used as negative controls, (B) NF-κB was activated in human mesenchymal stem cells (hMSCs) and osteoblast precursors (hOsteos) following 2 or 6 h of treatment with 100 ng/m TWEAK (Tw). Activation was measured using the TransAM NF-κB p65 activation assay system with cell lysates from normal and TNF-treated HeLa cells serving as negative and positive controls. The assays were carried out in triplicate and the data shown are representative of three independent experiments. (C) List of representative genes induced by TWEAK (100 ng/ml versus heat-inactivated TWEAK 100 ng/ml) in mesenchymal stem cells in low serum (LS: 0.2% FBS), moderate serum (MS: 2% FBS) and high serum (HS: 10% FBS). (D) List of some cell cycle-related genes induced by TWEAK (versus inactivated TWEAK) in mesenchymal stem cells cultured under low-serum conditions (0.2% FBS). Triplicate samples were analyzed for each condition and the fold changes were calculated using averages from triplicates. All fold changes reached statistical significance (P<0.01).

Article Snippet: These results therefore indicate that TWEAK may regulate cell fate decisions of progenitor cells. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Human mesenchymal progenitor cells are a novel target cell type for TWEAK. ( A ) Human primary mesenchymal stem cells, skeletal muscle myoblasts, preadipocytes, chondrocytes and osteoblast precursors (Cambrex) were cultured according to the manufacturer's protocols.

Techniques: Cell Culture, Staining, Binding Assay, Expressing, Activation Assay