Journal: Genes & Diseases

Article Title: Cancer-associated fibroblasts derived fibronectin extra domain A promotes sorafenib resistance in hepatocellular carcinoma cells by activating SHMT1

doi: 10.1016/j.gendis.2024.101330

Figure Lengend Snippet: CAFs promote sorafenib resistance by activating NF-κB in HCC cells. (A) Immunofluorescence analysis was performed to assess the expression of FAP and α-SMA on primary CAFs. Scale bars, 50 μm. (B) Co-culture with CAFs significantly reduced apoptosis of HepG2 and Huh7 upon sorafenib treatment. (blue bar: sorafenib-treated tumor cells cultured alone; red bar: sorafenib-treated tumor cells co-cultured with CAFs). Student's t -test of variance, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. (C) Enrichment score of the “I-κB kinase/NF-κB signaling” in sorafenib-resistant group versus sorafenib-sensitive group after sorafenib treatment, analyzed by Gene Set Enrichment Analysis based on the RNA-seq data (after performing log 2 transformation, normalization, and mean value calculation) obtained from the GEO database (GSE182593). (D) Western blot analyses of the protein level of P-p65 in the indicated HCC cells with different treatments. Results are representative of three experiments. (E) Inhibition of the NF-κB signaling pathway in HCC cells induced apoptosis significantly upon sorafenib treatment (blue bar: sorafenib-treated group; red bar: sorafenib and BAY11-7082-treated (100 μM) group). Student's t -test of variance, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. CAFs, cancer-associated fibroblasts; NF-κB, nuclear factor kappa B; HCC, hepatocellular carcinoma; FAP, fibroblast activation protein; α-SMA, alpha-smooth muscle actin.

Article Snippet: The cells on slides were fixed with 4% paraformaldehyde for 10 min and permeabilized in phosphate buffer saline for 20 min. Then, cells were blocked with goat serum at room temperature for 60 min and incubated with primary antibodies against FAP (fibroblast activation protein; R&D system, #FAB3715A, RRID: AB_2884010) (1:200) and α-SMA (alpha-smooth muscle actin; R&D system, #MAB1420, RRID: AB_262054) (1:200) for 2 h. Then, the cells on slides were reheated and incubated with the corresponding secondary antibody at 37 °C in the dark for 2 h. Nuclei were counter-stained with DAPI.

Techniques: Immunofluorescence, Expressing, Co-Culture Assay, Cell Culture, RNA Sequencing, Transformation Assay, Western Blot, Inhibition, Activation Assay

Journal: BMC Microbiology

Article Title: First human cell-based cultivation system for the syphilis spirochete Treponema pallidum

doi: 10.1186/s12866-026-04856-5

Figure Lengend Snippet: Human foreskin fibroblast cell lines support the growth of T. pallidum in vitro . A Human foreskin fibroblast cell lines (MoNa, HFF1, and HFFC) support the growth of T. pallidum in long-term, continuous, in vitro cultivation. While cultivation of strain SS14 was discontinued after 10 weeks, in vitro cultures of the DAL-1 strain have been ongoing for over one year. During this period, scheme of subcultures was optimized (up to 1:20). A booster passage (*), extending the standard 7-day subculture to a 14-day period with a cultivation medium exchange on day 7, was employed to enrich the treponemal culture during critical declines. In vitro cultivation was performed in triplicate (i.e., in three cultivation wells) and the mean values for each subculture are presented in graph. Source data from individual wells are shown in Supplementary Table . B Parallel cultivation for seven days ( n = 15) with defined DAL-1 inoculum (10 6 treponemes) validated the differences in growth support of individual foreskin cell lines. Treponemal growth on human foreskin fibroblast cells was slower compared to growth on rabbit epithelial Sf1Ep cells. Red bar, mean. C Human foreskin-based cultivation system supports the growth of various T. pallidum strains ( n = 8), from the Nichols-like as well as the SS14-like cluster. Note that human foreskin fibroblast cells were prepared as an equal mixture of three tested cell lines. Each T. pallidum strain was cultivated in a single in vitro well, representing a sole experimental replicate used for data acquisition

Article Snippet: Human foreskin fibroblasts HFF1 (SCRC-1041; ATCC) and HFFC (300715; Cytion) were purchased, while the third cell line (MoNa) was kindly provided by Dr. Vladimir Rotrekl (Masaryk University), and was originally obtained from the National Tissue Centre (Czech Republic).

Techniques: In Vitro