Journal: Lipids in Health and Disease

Article Title: Active compound chrysophanol of Cassia tora seeds suppresses heat-induced lipogenesis via inactivation of JNK/p38 MAPK signaling in human sebocytes

doi: 10.1186/s12944-019-1072-x

Figure Lengend Snippet: PPARγ and FAS are upregulated by heat in human sebocytes. Human sebocytes were serum-starved for 6 h and then incubated for 30 min at 41 °C or 44 °C. Fresh culture medium was added and the cells were cultured. a Cell viability was assessed by MTT assay. b , c PPARγ, FAS and target gene of PPARγ protein levels were analyzed by western blotting, with actin used for normalization. d Intracellular lipids in human sebocytes exposed to heat were assessed by Nile Red staining. Scale bar = 20 μm. Quantitation of lipid levels in human sebocytes exposed to heat, calculated as percentage of the value in cells incubated at 37 °C. Data represent the mean ± SE ( n = 3). * P < 0.05, (Student’s t test)

Article Snippet: Human sebocytes were purchased from ZenBio (Research Triangle Park, NC, USA) and cultured in Sebomed medium (Biochrom ag , Berlin, Germany) supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, UT, USA) and 5 ng ml − 1 recombinant human epidermal growth factor (Biochrom ag , Berlin, Germany).

Techniques: Incubation, Cell Culture, MTT Assay, Western Blot, Staining, Quantitation Assay

Journal: Lipids in Health and Disease

Article Title: Active compound chrysophanol of Cassia tora seeds suppresses heat-induced lipogenesis via inactivation of JNK/p38 MAPK signaling in human sebocytes

doi: 10.1186/s12944-019-1072-x

Figure Lengend Snippet: Inhibition of lipid synthesis by CTSE and CP in heat-treated sebocytes. Human sebocytes were serum-starved for 6 h and then incubated for 30 min at 44 °C. Fresh culture medium was added with various concentrations of CTSE (0–100 ppm) or CP (0–100 μM), followed by incubation for 48 h. All experiments included non-heat-shocked and untreated sebocytes as a control. a Cells were treated with various concentrations of CTSE (0–100 ppm) and CP (0–100 μM), and cell survival was analyzed using the MTT reduction assay. Data represent the mean ± SD of triplicate samples expressed as a percentage of the control. b , c Cell lysates were analyzed by western blotting for PPARγ and FAS with actin used for normalization. d Levels of intracellular lipids assessed by Nile Red staining. Scale bar = 20 μm. Lipid levels in heat-shocked and CTSE or CP-treated human sebocytes, calculated as a percentage of the value in cells incubated at 37 °C. Data represent the mean ± SE ( n = 3). * P < 0.05 vs. cells incubated at 37 °C; † P < 0.05, vs. cells incubated at 44 °C, (Student’s t test)

Article Snippet: Human sebocytes were purchased from ZenBio (Research Triangle Park, NC, USA) and cultured in Sebomed medium (Biochrom ag , Berlin, Germany) supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, UT, USA) and 5 ng ml − 1 recombinant human epidermal growth factor (Biochrom ag , Berlin, Germany).

Techniques: Inhibition, Incubation, Control, MTT Reduction Assay, Western Blot, Staining

Journal: Lipids in Health and Disease

Article Title: Active compound chrysophanol of Cassia tora seeds suppresses heat-induced lipogenesis via inactivation of JNK/p38 MAPK signaling in human sebocytes

doi: 10.1186/s12944-019-1072-x

Figure Lengend Snippet: Heat induces lipid synthesis in human sebocytes via activation of JNK/p38 signaling. Human sebocytes were starved for 6 h and then incubated for 30 min at 44 °C. Fresh culture medium was added and incubated. a Whole cell lysates were prepared in a time-dependent manner and analyzed by western blotting for JNK and phospho-JNK, and p38 and phospho-p38. b Human sebocytes were incubated for 30 min at 44 °C and treated with various concentrations of CP (0–100 μM), followed by incubation for 48 h

Article Snippet: Human sebocytes were purchased from ZenBio (Research Triangle Park, NC, USA) and cultured in Sebomed medium (Biochrom ag , Berlin, Germany) supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, UT, USA) and 5 ng ml − 1 recombinant human epidermal growth factor (Biochrom ag , Berlin, Germany).

Techniques: Activation Assay, Incubation, Western Blot

Journal: Lipids in Health and Disease

Article Title: Active compound chrysophanol of Cassia tora seeds suppresses heat-induced lipogenesis via inactivation of JNK/p38 MAPK signaling in human sebocytes

doi: 10.1186/s12944-019-1072-x

Figure Lengend Snippet: Reduction of pro-inflammatory cytokine levels by CP treatment of heat-shocked human sebocytes. Human sebocytes were serum-starved for 6 h and then incubated for 30 min at 44 °C. Fresh culture medium was added with various concentrations of CP (0–100 μM), followed by incubation for 48 h. a Cytokine levels were detected with an array that included antibodies against 42 different cytokines. Neg, negative control; Pos, positive control. b Three pro-inflammatory cytokines were upregulated by heat treatment (red squares); this effect was mitigated by treatment with CP. c Densitometry analysis of cytokine levels. d Pro-IL-1β protein levels in the whole cell extract and mature IL-1β protein level in the culture supernatant were determined by western blotting, with actin used for normalization of the whole cell lysate. e Mature IL-1β in the culture supernatant was detected by ELISA. Data represent the mean ± SEM ( n = 3). * P < 0.05 vs. cells incubated at 37 °C; † P < 0.05 vs. cells incubated at 44 °C, (Student’s t test)

Article Snippet: Human sebocytes were purchased from ZenBio (Research Triangle Park, NC, USA) and cultured in Sebomed medium (Biochrom ag , Berlin, Germany) supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, UT, USA) and 5 ng ml − 1 recombinant human epidermal growth factor (Biochrom ag , Berlin, Germany).

Techniques: Incubation, Negative Control, Positive Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Lipids in Health and Disease

Article Title: Active compound chrysophanol of Cassia tora seeds suppresses heat-induced lipogenesis via inactivation of JNK/p38 MAPK signaling in human sebocytes

doi: 10.1186/s12944-019-1072-x

Figure Lengend Snippet: Expression of NF-kB signaling by heat treatment in human sebocytes. Human sebocytes were starved for 6 h and then incubated for 30 min at 44 °C. Fresh culture medium was added and incubated. a Whole cell lysates were prepared in a time-dependent manner and analyzed by western blotting for NFκB and phospho- NFκB, and IκB α and phospho- IκB α. b Human sebocytes were incubated for 30 min at 44 °C and treated with various concentrations of CP (0–100 μM), followed by incubation for 48 h

Article Snippet: Human sebocytes were purchased from ZenBio (Research Triangle Park, NC, USA) and cultured in Sebomed medium (Biochrom ag , Berlin, Germany) supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, UT, USA) and 5 ng ml − 1 recombinant human epidermal growth factor (Biochrom ag , Berlin, Germany).

Techniques: Expressing, Incubation, Western Blot