Journal: Oncogenesis

Article Title: Altering MYC phosphorylation in the epidermis increases the stem cell population and contributes to the development, progression, and metastasis of squamous cell carcinoma

doi: 10.1038/s41389-020-00261-3

Figure Lengend Snippet: a , b TUNEL staining ( a ) and quantitation ( b ) to assess apoptosis in the epidermis and hair follicle of mice 72 h post DMBA/TPA one-time treatment. Scale bar = 50 µm. Shown is mean and SD of 5 mice per genotype. p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, *** p < 0.001. c , d BrdU staining ( c ) and quantitation ( d ) to assess proliferation in the epidermis and hair follicle of mice 48 h post DMBA/TPA one-time treatment. Scale bar = 100 µm. Shown is mean and SD of 3 mice per genotype. p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, ns = not significant. e , f BrdU staining ( e ) and quantitation ( f ) in the epidermis adjacent to the skin lesions after 8 weeks of DMBA/TPA treatment of mice from each genotype. Scale bar = 50 µm. Shown is mean and SD of 5 mice per genotype. p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, ** p < 0.01, *** p < 0.001. g , h TUNEL staining ( g ) and quantitation ( h ) to assess apoptosis in the epidermis and hair follicle of mice 8 weeks post DMBA/TPA treatment. Scale bar = 100 µm. Shown is mean and SD of 3 mice per genotype. p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, ns = not significant.

Article Snippet: Briefly, the paraffin slides were de-paraffinized, incubated with TdT enzyme at 37 °C for 1 h, washed in TACS 2 TdT stop buffer for 10 min, incubated with streptavidin-HRP for 30 min, and counterstained with methanol green for 10 min. For proliferation analysis, a BrdU cell proliferation Assay Kit (BioVision, Cat. K306), according to the manufacturer’s protocol.

Techniques: TUNEL Assay, Staining, Quantitation Assay, Comparison, BrdU Staining

Journal: Oncogenesis

Article Title: Altering MYC phosphorylation in the epidermis increases the stem cell population and contributes to the development, progression, and metastasis of squamous cell carcinoma

doi: 10.1038/s41389-020-00261-3

Figure Lengend Snippet: a , b Long-term label-retaining cell analysis in control, MYC WT , or MYC T58A mice. Retention of BrdU was assessed 75 days after injection through immunofluorescent staining for BrdU (red) shown in ( a ) and quantification of percent of cells positive for BrdU shown in ( b ). p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, *** p < 0.001. c Immunofluorescence images of skin from the long-term label-retaining analysis stained with anti-CD34 (red). White arrows indicate positive cells in hair follicles. DAPI (blue) is nuclear counterstain. d Quantitation of CD34 staining from (c). Significance was determined by one-way ANOVA, followed by Tukey’s multiple comparison test, ** p < 0.01, *** p < 0.001. e qRT-PCR for Lin28B expression in skin of mice used in long-term label-retaining analysis (normal skin). p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, * p < 0.05, ** p < 0.01. f Immunohistochemistry for CD34 staining in hair follicles from the hyperplastic skin of control, MYC WT , or MYC T58A mice 4 months after DMBA/TPA treatment. Scale bar = 50 µm. Insets are shown below. Red arrowheads indicate CD34 positive cells. Nonspecific staining in the sebaceous gland is indicated by asterisks. g Quantification of CD34 staining from mice as in ( f ). Shown is mean and SD of 5 mice per genotype. p -value is from a one-way ANOVA, followed by Tukey’s multiple comparison test, * p < 0.05, *** p < 0.001. h qRT-PCR for expression of stem cell marker or signaling pathway genes, as indicated, in hyperplastic epidermis from DMBA/TPA-treated control, MYC WT , or MYC T58A mice. p -value is from a two-tailed Welch’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Briefly, the paraffin slides were de-paraffinized, incubated with TdT enzyme at 37 °C for 1 h, washed in TACS 2 TdT stop buffer for 10 min, incubated with streptavidin-HRP for 30 min, and counterstained with methanol green for 10 min. For proliferation analysis, a BrdU cell proliferation Assay Kit (BioVision, Cat. K306), according to the manufacturer’s protocol.

Techniques: Cell Analysis, Control, Injection, Staining, Comparison, Immunofluorescence, Quantitation Assay, Quantitative RT-PCR, Expressing, Immunohistochemistry, Marker, Two Tailed Test

Journal: Advanced Science

Article Title: Palmitoyltransferase ZDHHC3 Aggravates Nonalcoholic Steatohepatitis by Targeting S ‐Palmitoylated IRHOM2

doi: 10.1002/advs.202302130

Figure Lengend Snippet: Hepatocyte‐specific loss of Zdhhc3 protects against HFHC‐induced NASH pathogenesis. a‐g) Records for the body weight (a), liver weight and the ratio of liver weight/body weight (%) (LW/BW) (b), fasting blood glucose levels (c), fasting insulin levels (d), HOMA‐IR index (e), glucose tolerance test (GTT) (f) and liver TG, NEFA and TC contents (g) of the HFHC‐fed Zdhhc3 ‐Flox mice and Zdhhc3 ‐HepKO mice; NCD diet‐fed mice were treated as corresponding control ( n = 10 mice per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). h) Pearson analysis indicating the correlations between fasting insulin, ratio of liver weight/body weight (%), and liver TG contents in indicated groups ( n = 10 per parameter; P < 0.001 for all of these correlations). i‐k) Representative pictures of H&E staining, oil red O staining, histological NAS score (i) changes, sirius red staining, masson staining (j), and F4/80 and CD11b positive cells expression (k) in indicated groups (magnification, 100× for H&E staining, oil red O staining, masson staining and sirius red staining; magnification, 200× for F4/80 and CD11b staining; n = 10 images per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). l, Records for inflammation‐related cytokines profiles including IL‐6, TNF‐α, IL‐1β, IL‐18 and CCL2 in serum from HFHC‐fed HepKO or Flox mice ( n = 10 mice per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). m, Records for liver function‐related indicators including ALT, AST, AKP, and GGT in serum from HFHC‐fed HepKO or Flox mice ( n = 10 mice per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). Data are expressed as mean ± SEM. The relevant experiments presented in this part were performed independently at least three times. Significance is determined by two‐way analysis of variance (ANOVA) followed by multiple comparisons test analysis (a‐g) or 2‐sided Student's t ‐test (i‐m). The P ‐value < 0.05 was considered as significant difference.

Article Snippet: The TNF‐α (Cat: ab100747), IL‐1β (Cat: ab197742), IL‐6 (Cat: ab222503), CCL2 (Cat: ab208979), IL‐18 (Cat: ab216165) and insulin (Cat: ab285341) ELISA kits were purchased from Abcam.

Techniques: Control, Staining, Expressing