Journal: Clinical Cancer Research

Article Title: Anti-EGFR Antibody–Drug Conjugate Carrying an Inhibitor Targeting CDK Restricts Triple-Negative Breast Cancer Growth

doi: 10.1158/1078-0432.CCR-23-3110

Figure Lengend Snippet: Stochastic conjugation of cetuximab to CDK inhibitor and ADC internalization in live breast cancer cells. A, Flow cytometric evaluation of surface EGFR expression (TNBC: MDA-MB-468, HCC1143, HCC1806, MDA-MB-231, HCC1937, SUM149, and CAL51; HER2+: SKBR3; ER+: MCF7, T47D; nontumorigenic epithelial cell model: MCF10A; immune cell model: human B lymphocytes RPMI8866, RPMI8226, and monocytic cell line U937; human primary melanocyte: melanocyte). B, EGFR mRNA expression from the Cancer Cell Line Encyclopedia database showed a positive correlation with surface EGFR measured by flow cytometry in A (Spearman’s rank coefficient, r = 0.723). A high level of correlation was found between EGFR and cyclin E ( r = 0.738), but not with cyclin A or CDK2. Nonsignificant P values are marked as NS. C, Top, Schematic diagram of stochastic ADC conjugation by antibody reduction with TCEP and then conjugation to SNS-032 via MC-Val–Ala-PAB. Middle, HIC analysis confirmed an average DAR of 4.4. Bottom, SEC trace indicates negligible ADC aggregation and minimal free linker–payload (less than 0.8%). D, Surface plasmon resonance analysis demonstrated similar binding affinity ( K D ) for cetuximab (0.73 nmol/L) and ADC (1.28 nmol/L). Isotype IgG1 and isotype ADC showed no measurable binding. E, Monitoring internalization of Fabfluor-pH-labeled cetuximab, ADC, or isotype control (10 nmol/L) by Incucyte live-cell imaging. Phase and red fluorescence time-course images were captured for 24 hours. Images of internalized antibody display in cytosolic, low pH lysosomal vesicle-associated red fluorescence in cells. Scale bar, 0.2 mm. F, Cells were seeded in Matrigel for 5 days, allowing the formation of spheroids. Fabfluor-pH-labeled antibodies or ADC (10 nmol/L) were introduced in the Matrigel and showed rapid internalization in EGFR-high MDA-MB-468 and MDA-MB-231, whereas EGFR-low CAL51 displayed little red fluorescence signals. A low level of internalization was observed for isotype or isotype-ADC controls. Scale bar, 0.5 mm. P values determined by two-tailed unpaired t test of three independent experiments compared with isotype control.

Article Snippet: Human primary epidermal melanocytes were cultured in Melanocyte Growth Medium (Cell Applications Inc.).

Techniques: Conjugation Assay, Expressing, Flow Cytometry, SPR Assay, Binding Assay, Labeling, Control, Live Cell Imaging, Fluorescence, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Rewiring melanoma cell fate: TRPM8 modulators trigger apoptosis and boost NK cell cytotoxicity

doi: 10.1038/s41419-026-08469-8

Figure Lengend Snippet: A Representative Western blot showing TRPM8 protein expression in the indicated cell lines. Tubulin was used as loading control. Viability of human melanocytes ( B ) and human dermal fibroblasts ( C ) untreated or treated with compounds 4 and 9 at the concentrations indicated in the legends on the right. Absorbance values from WST-1 assays at 24, 48, and 72 h are shown. Data are presented as mean ± SD of three independent experiments. n.s . indicates not significant. Representative Live/Dead assay images of human melanocytes ( D ) and human dermal fibroblasts ( E ) treated for 24 h with compounds 4 and 9 (1 or 10 μM). Viable cells are shown in green (acridine orange; total cells), while dead cells are shown in red (propidium iodide; dead cells). Overlay images are shown. Scale bar, 100 μm. Quantification of cell death is displayed to the right of each overlay image. The percentage of dead cells was calculated as: (red-stained dead cells/green-stained total cells) × 100.

Article Snippet: The human epidermal Melanocytes ( P10853 , Innoprot, Bizkaia, Spain) were cultured in Melanocyte Medium (MeIM; Innoprot) supplemented with 5% FBS (Gibco), 1% Melanocyte Growth Supplement (MelSG; Innoprot) and 1% penicillin-streptomycin (Gibco).

Techniques: Western Blot, Expressing, Control, Live Dead Assay, Staining

Journal: Nutrients

Article Title: Evaluation of the Relationship Between Orexin A, Peptide YY, AgRP, and POMC Levels and Sleep Disorders in Children with Malnutrition

doi: 10.3390/nu18030377

Figure Lengend Snippet: Box plots showing the distribution of the orexin A, proopiomelanocortin (POMC), agouti-related protein (AgRP), and peptide yy (PYY) levels in the malnourished children and typically developing (TD) healthy controls. The Mann–Whitney U test was used to compare the peptide levels between the two groups.

Article Snippet: Human POMC, orexin A, AgRP, and PYY enzyme-linked immunosorbent assay (ELISA) kits (Shanghai Korain Biotech Co., Ltd., Shanghai, China) were used to measure the plasma peptide levels.

Techniques: MANN-WHITNEY

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Effects of tea polyphenols on UVA-induced melanogenesis via inhibition of α-MSH-MC1R signalling pathway

doi: 10.5114/ada.2022.115890

Figure Lengend Snippet: Effects of TPS on the expression of α-MSH. Representative chart trace of α-MSH inhibition by 5, 10, and 15 μg/ml of the TPS on HaCaT cells ( A ) and HEM cells ( B ) with UVA exposure (15 J/cm 2 ) or not by ELISA. Error bars show means ± SEMs. * P < 0.05, ** p < 0.01, *** p < 0.001 versus non-treated cells, # p < 0.05, ## p < 0.01, ### p < 0.001 versus UVA-treated cells

Article Snippet: The following material was purchased from the following manufacturers: tea polyphenols (98% purity), Chinese Academy of Agricultural Sciences, Tea Research Institute; 10% foetal bovine serum (FBS), DMEM and DMSO, Gibco/BRL (Grand Island, NY, USA); Bovine serum albumin (BSA), levodopa (L-DOPA) and nonapeptide-1 acetate salt (N-1A), MedChemExpress (Shanghai, China); and α-MSH ELISA Kit, CUSABIO (Wuhan, China).

Techniques: Expressing, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Effects of tea polyphenols on UVA-induced melanogenesis via inhibition of α-MSH-MC1R signalling pathway

doi: 10.5114/ada.2022.115890

Figure Lengend Snippet: TPS inhibited melanogenesis through suppressing the α-MSH-MC1R signalling pathway in HaCaT cells and HEM cells. A, B – HEM was pretreated or not with UVA exposure (15 J/cm 2 ) before TPS (10 μg/ml) and N-1A (20 μm) were applied. Melanin contents and the expression of tyrosinase were measured as described in methods. C – HaCaT cells were treated with TPS (10 μg/ml) and/or N-1A (20 μm) in the presence or absence of UVA exposure (15 J/cm 2 ). ELISA was then applied to detect the expression of α-MSH. D – HEM cells were treated with TPS (10 μg/ml) and N-1A (20 μm) in the presence or absence of UVA exposure (15 J/cm 2 ). ELISA was then applied to detect the expression of α-MSH. * P < 0.05, ** p < 0.01, *** p < 0.001 versus non-treated cells, # p < 0.05, ## p < 0.01, ### p < 0.001 versus UVA-treated cells, a p < 0.05, aa p < 0.01, aaa p < 0.001, ns p > 0.05

Article Snippet: The following material was purchased from the following manufacturers: tea polyphenols (98% purity), Chinese Academy of Agricultural Sciences, Tea Research Institute; 10% foetal bovine serum (FBS), DMEM and DMSO, Gibco/BRL (Grand Island, NY, USA); Bovine serum albumin (BSA), levodopa (L-DOPA) and nonapeptide-1 acetate salt (N-1A), MedChemExpress (Shanghai, China); and α-MSH ELISA Kit, CUSABIO (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay