Journal: bioRxiv

Article Title: Heparan sulfate promotes autoactivation of pro-Cathepsin K by destabilizing the propeptide-catalytic domain interaction

doi: 10.64898/2026.01.18.700217

Figure Lengend Snippet: Mouse femur section was co-stained with anti-CtsK (green), HS20 (purple) and anti-LAMP2 (red) and visualized with anti-rabbit Alexa488, anti-human Alexa594, and anti-rat Alexa647 secondary antibodies, respectively. Area shown is the cortical bone. The approximate boundary of the bone is shown with white dashed line. A blood vessel within the bone is indicated with yellow dashed line. Red blood cells in the vessel shown strong green autofluorescence. A closeup view of the large osteoclast is shown in the right bottom panel. Another osteoclast (likely only the tip of the osteoclast) is indicated with a yellow arrow in the merged image. Nuclei of several osteocytes are indicated with white arrow head. Images representative of three separate experiments.

Article Snippet: Following deparaffinization and citric acid-based antigen retrieval (pH7), sectioned were blocked and incubated overnight at 4 °C with the following primary antibodies: 0.2 μg/ml rabbit anti-mCtsK polyclonal antibody (described previously) , 1ug/ml human anti-HS mAb (HS20, from Bio X cell) , and 1ug/ml Rat anti-mouse Lamp2 (GL2A7, Developmental Studies Hybridoma Bank).

Techniques: Staining

Journal: Microbiology Spectrum

Article Title: Sex-specific transcriptional regulation in lung macrophages during sub-acute Mycobacterium tuberculosis infection

doi: 10.1128/spectrum.01790-25

Figure Lengend Snippet: Markers of inflammation, cell survival, and autophagy exhibit sex-specific regulation in murine lungs during sub-acute Mtb infection. ( a ) Immunoblot analysis of TNF-α, IFN-γ, IL-6, phosphorylated AKT (p-AKT), and ANNEXIN V in lung protein lysates from male and female mice. GDI or total protein serves as the loading control. Representative blots highlight variations between sexes. ( b ) Bar graph values were derived from densitometric quantification of protein bands normalized to GDI or total protein, showing sex-related differences in inflammatory and cell survival signaling pathway. Data are presented as mean ± SEM. ( c ) qPCR analysis of autophagy-related genes ( Lamp2 , Ulk1 , Atg7 , Becn1 , Lc3 , and Tubb1 ) in lung tissue from male and female mice. Gene expression levels are normalized to 18S rRNA and presented as fold change. The error bars represent the standard error of the mean. Statistical significance was calculated using t -test. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 between indicated groups.

Article Snippet: RNA was reverse-transcribed from 100 ng of total RNA using iScript cDNA Synthesis Kit (#1708891, Bio Rad), and mRNA levels of IgHg1 , IL-1b (Interleukin 1b), Ctsc (Cathepsin C), Arg1 (Arginase), Cacna1c (Calcium Channel Alpha 1c), Lamp2 (Lysosome-associated membrane protein 2), Ulk1 (Unc-51 like autophagy activating kinase 1), Tubb1 (Tubulin, Beta 1 Class VI), Adra1 (Adrenergic Receptor Alpha 1), and Col9a1 (Collagen type IX alpha 1 chain) were analyzed by qPCR using iTaq universal SYBR green super mix (#1725124, Bio Rad) and normalized to 18S levels as previously demonstrated ( ).

Techniques: Infection, Western Blot, Control, Derivative Assay, Gene Expression

Journal: Microbiology Spectrum

Article Title: Sex-specific transcriptional regulation in lung macrophages during sub-acute Mycobacterium tuberculosis infection

doi: 10.1128/spectrum.01790-25

Figure Lengend Snippet: Differentially expressed genes in lung macrophages isolated from Mtb -infected male and female. ( a ) KEGG pathway enrichment analysis showing upregulated signaling pathways enriched in the lung macrophages from the female mice in response to Mtb infection. ( b ) KEGG pathway enrichment analysis of upregulated genes in lung macrophages from the male mice, illustrating pathways that are preferentially activated in the male mice in response to infection. ( c ) qPCR validation of RNA-seq data in lung macrophages from Mtb -infected mice. Quantitative PCR (qPCR) analysis confirming the differential expression of genes upregulated in male macrophages, including Ighg1 , Il1b , Ctsc , and Arg1 , showing higher expression levels in males compared to females identified by RNA sequencing. ( d ) Genes upregulated in female macrophages, including Cacna1c , Lamp2 , Ulk1 , Tubb1 , Adra1a, and Col9, exhibiting higher expression in females. Gene expression levels are normalized to 18S rRNA and presented as fold change. Data are shown as mean ± SEM. Statistical significance was determined using a t -test between infected male and female mice. * P < 0.05, ** P < 0.01 between indicated groups.

Article Snippet: RNA was reverse-transcribed from 100 ng of total RNA using iScript cDNA Synthesis Kit (#1708891, Bio Rad), and mRNA levels of IgHg1 , IL-1b (Interleukin 1b), Ctsc (Cathepsin C), Arg1 (Arginase), Cacna1c (Calcium Channel Alpha 1c), Lamp2 (Lysosome-associated membrane protein 2), Ulk1 (Unc-51 like autophagy activating kinase 1), Tubb1 (Tubulin, Beta 1 Class VI), Adra1 (Adrenergic Receptor Alpha 1), and Col9a1 (Collagen type IX alpha 1 chain) were analyzed by qPCR using iTaq universal SYBR green super mix (#1725124, Bio Rad) and normalized to 18S levels as previously demonstrated ( ).

Techniques: Isolation, Infection, Protein-Protein interactions, Biomarker Discovery, RNA Sequencing, Real-time Polymerase Chain Reaction, Quantitative Proteomics, Expressing, Gene Expression