Journal: bioRxiv

Article Title: HTRA3 protease-chaperone stabilizes cathepsin B for mitochondrial POLG1 depletion in human cell ageing

doi: 10.1101/2025.07.21.647696

Figure Lengend Snippet: (A) WB of CSB, HTRA3 (left panel), and CTSB, and p21 (right panel) in BJ fibroblasts untreated and 10 days post-irradiation using ß-tubulin and GAPDH, respectively, as loading control. (B) Scheme showing the two hypotheses for the mechanism leading to senescence-related HTRA3 and CTSB overexpression. (C) Immunoblots of p21, CSB, HTRA3, CTSB, and POLG1 in BJ-5ta hTERT WT fibroblasts and 2 CRISPR-edited CSB knocked-out clones (BJ-5ta csb-/- hTERT #1 and BJ-5ta csb-/- hTERT #2). Frames show samples on the same blot; each displaying the respective GAPDH used as loading control. (D) Scheme indicating normally blocked senescence in BJ-5ta hTERT cells and activation of senescence upon irradiation at 10 Gy. WB analysis of (E) CSB, CTSB, p21, and (F) HTRA3 three days post-irradiation in BJ-5ta hTERT WT fibroblasts and BJ-5ta hTERT CSB knocked-out fibroblasts (β-tubulin was used as loading control). RT-qPCR of ( G ) p21 Waf1 and ( H ) HTRA3 and ( I ) CTSB in the same experiment and cells schematized in panel D. Samples on the same blot are framed; each frame displays the respective GAPDH or β-tubulin used as loading control. RT-qPCR: n=3 independent experiments; mean ± SD. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001; based on unpaired t test comparisons to respective non-irradiated control.

Article Snippet: Normal female human foetal lung IMR-90 fibroblasts (ATCC; CCL-186) and BJ skin fibroblasts (ATCC; CRL-2522) were cultured in minimum essential medium (MEM, Gibco) supplemented with 2 mM L-glutamin (GlutMAX), 10% FBS, 1% Penicillin–streptomycin, 1% nonessential amino acids (Gibco) and 1% sodium pyruvate (Gibco) BJ-5ta hTERT immortalized human fibroblasts (ATCC; CCL-186) cultured in a 4:1 mixture of DMEM and Medium 199 (Gibco) supplemented with 10% FBS and1% Penicillin–streptomycin.

Techniques: Irradiation, Control, Over Expression, Western Blot, CRISPR, Clone Assay, Activation Assay, Quantitative RT-PCR

Journal: Cell Death Discovery

Article Title: New AKT-dependent mechanisms of anti-COVID-19 action of high-CBD Cannabis sativa extracts

doi: 10.1038/s41420-022-00876-y

Figure Lengend Snippet: A Western blot analysis of ACE2 and TMPRSS2 in WI-38 cells treated with the indicated most abundant compounds in extract #1, with the same concentration found in the extract; relative densitometry was measured using ImageJ. B Western blot analysis of ACE2 and TMPRSS2 in BJ-5ta cells treated with the indicated most abundant compounds in extract #7, with the same concentration found in the extract; relative densitometry was measured using ImageJ. C Western blot analysis of ACE2 and TMPRSS2 in WI-38 cells treated with either CBD or PTWT2.2 individually or in combination; relative densitometry was measured using ImageJ. D Western blot analysis of ACE2 and TMPRSS2 in WI-38 cells treated with either CBD or PTWT1.2 individually or in combination; relative densitometry was measured using ImageJ. PTWT represents terpene. * indicates p < 0.05; ** indicates p < 0.01.

Article Snippet: Human normal foreskin fibroblasts (BJ-5ta), purchased from American Type Culture Collection (ATCC, Manassas, USA), were cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum (FBS).

Techniques: Western Blot, Concentration Assay

Journal: PLoS Pathogens

Article Title: An apical protein, Pcr2, is required for persistent movement by the human parasite Toxoplasma gondii

doi: 10.1371/journal.ppat.1010776

Figure Lengend Snippet: A. Images selected from time-lapse experiments of intracellular RH ΔhxΔku80 ( WT ), mEmeraldFP-Pcr2 knock-in ( mE-Pcr2 KI ), and knockout ( Δpcr2 ) treated with 5 μM A23187 (also see ). The cell-impermeant DNA-binding dye, DAPI, was added to the medium to monitor the permeabilization of the host cell. Δpcr2 parasites are able to secrete effectors that lyse the host cell upon A23187 treatment, indicated by DAPI entering the host cell nucleus and binding to DNA, as well as by the dramatic change in the morphology of the host cell (see and Videos). Insets are DAPI images of the nuclear region of the host cell shown at 0.5X. Brackets in the mE-Pcr2 KI panels indicate the host cell nucleus included in the insets. Contrast was adjusted so that the DAPI labeling at the rim of the nucleus is easily visible. The nuclei of uninfected fibroblasts (marked by dashed circles) remained unlabeled by DAPI ~19 min after A23187 treatment as shown in the larger field of view images in the right-hand column. B. Projections of deconvolved wide-field fluorescence images of intracellular WT, mE-Pcr2 KI, Δpcr2 , and complemented (Comp) parasites labeled with a mouse anti-MIC2 (red), a rat anti-GAP45 (cyan) and corresponding secondary antibodies. C. Western blots of the secreted (supernatant, S) and unsecreted (pellet, P) fractions of WT, mE-Pcr2 KI, Δpcr2 , and complemented (Comp) parasites after A23187 or BAPTA-AM (a calcium chelator; negative control) treatment. The blots were probed by antibodies against MIC2 and GRA8. M: molecular weight markers, the masses of which are indicated in kDa by the numbers on the left. D. Levels of MIC2 in the secreted fractions relative to that from the wild-type in 3 independent biological replicates. For each sample, the MIC2 secretion upon A23187 stimulation is normalized against GRA8 in the pellet from the same sample. Error bars: standard error.

Article Snippet: Confluent human foreskin fibroblasts (HFFs; ATCC# SCRC-1041, and HFF_hTERT; ATCC# CRL-4001) monolayers in Dulbecco’s Modified Eagle’s Medium (DMEM, VWR, 45000–316), supplemented with 1% (v/v) heat-inactivated Cosmic calf serum (Hyclone, SH30087.3) and Glutamax (Life Technologies-Gibco, 35050061) were used to maintain parasite cultures.

Techniques: Knock-In, Knock-Out, Binding Assay, Labeling, Fluorescence, Western Blot, Negative Control, Molecular Weight

Journal: ACS Applied Materials & Interfaces

Article Title: Hybrid Tellurium–Lignin Nanoparticles with Enhanced Antibacterial Properties

doi: 10.1021/acsami.0c22301

Figure Lengend Snippet: (a) Cell viability (%) of human keratinocytes and fibroblasts treated with different concentrations of TeLigNPs (ppm of Te) for 24 h assessed by the AlamarBlue assay. Results are reported as mean values ± SD ( n = 3). (b) Live/dead assay of human keratinocytes and fibroblasts treated with different concentrations of TeLigNPs (ppm of Te). The assay stains the live cells in green and the dead ones in red.

Article Snippet: Bacterial strains S. aureus (ATCC 25923), E. coli (ATCC 25922), and P. aeruginosa (ATCC 10145), human fibroblast cells (ATCC-CRL-4001, BJ-5ta), and human keratinocyte cells (HaCaT cell line) were obtained from the American Type Culture Collection (ATCC LGC Standards, Spain).

Techniques: Alamar Blue Assay, Live Dead Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Hybrid Tellurium–Lignin Nanoparticles with Enhanced Antibacterial Properties

doi: 10.1021/acsami.0c22301

Figure Lengend Snippet: ROS generation assessment using the fluorescent probe H 2 DCFDA after incubation of (a) bacteria ( S. aureus , E. coli , and P. aeruginosa ) and (b) human keratinocytes and fibroblasts with TeLigNPs. Results are reported as mean values ± SD ( n = 3).

Article Snippet: Bacterial strains S. aureus (ATCC 25923), E. coli (ATCC 25922), and P. aeruginosa (ATCC 10145), human fibroblast cells (ATCC-CRL-4001, BJ-5ta), and human keratinocyte cells (HaCaT cell line) were obtained from the American Type Culture Collection (ATCC LGC Standards, Spain).

Techniques: Incubation, Bacteria