human normal primary non immortalized dermal fibroblasts hdfa  (ATCC)

Journal: Frontiers in Immunology

Article Title: Identification and histological validation of autophagy-related core genes ADRB2 and PLK2 in keloids, with integrated immune infiltration analysis

doi: 10.3389/fimmu.2026.1724230

Figure Lengend Snippet: Functional validation of ADRB2 and PLK2 in keloid fibroblasts via autophagy modulation. (A) Western blot analysis comparing NFs and KFs shows increased protein expression of ADRB2, PLK2, COL1, COL3, LC3-II/LC3-I ratio, and p62 in KFs, with GAPDH used as a loading control. (B) Densitometric quantification of Western blot bands demonstrates significantly higher levels of ADRB2, PLK2, COL1, COL3, LC3-II/LC3-I, and p62 in KFs compared with NFs (*p < 0.05, **p < 0.01, ***p < 0.001). (C) Representative fluorescence images of MDC staining indicate reduced MDC-positive puncta in KFs compared with NFs, while EBSS treatment increases and CQ treatment decreases MDC-labeled autophagic structures. (D) Western blot analysis of KFs treated with EBSS or CQ shows that autophagy modulation alters fibrotic markers (COL1 and COL3), autophagy-related proteins (LC3-I/LC3-II and p62), and ADRB2 expression, whereas PLK2 levels remain relatively stable across treatments. (E) Quantitative analysis confirms that EBSS-induced autophagy activation reduces COL1, COL3, p62, and ADRB2 expression, while CQ-mediated autophagy inhibition results in increased levels of these proteins, with no significant change observed in PLK2 expression (ns, not significant; *p < 0.05, **p < 0.01).

Article Snippet: Normal human dermal fibroblasts (NFs; CRL-4066) and keloid-derived fibroblasts (KFs; CRL-1762) were obtained from ATCC (USA).

Techniques: Functional Assay, Biomarker Discovery, Western Blot, Expressing, Control, Fluorescence, Staining, Labeling, Activation Assay, Inhibition