Journal: bioRxiv

Article Title: Mechanistic dissection of Candidatus Liberibacter Triggered Chronic Immune Disease

doi: 10.1101/2025.05.21.654963

Figure Lengend Snippet: ( a ) Left: Tomato plants of the indicated genotypes were inoculated with Candidatus Liberibacter psyllaurous (Lpsy). Quantitative PCR (qPCR) was performed 21 days post-inoculation (DPI) to assess bacterial titers (Ct values). The photograph was taken at 60 DPI. Right: qPCR analysis was performed 6 weeks post-inoculation, and the corresponding image was captured at 7 weeks post-inoculation. ( b ) Trypan blue staining of tomato leaves from healthy and Lpsy-infected plants, highlighting cell death in infected tissues. ( c ) Cell death in tomato leaves of the indicated genotypes, measured by ion leakage assay. Ion leakage was calculated as the percentage of initial ion conductivity relative to total conductivity after boiling. ( d ) Reactive oxygen species (ROS) detection in H₂DCFDA-stained tomato leaves, visualized using a PerkinElmer IVIS Lumina S5 imaging system. Red fluorescence indicates ROS accumulation. ( e ) Quantification of leaf starch content in healthy (control) and Lpsy-infected tomato plants. Total starch levels were normalized to leaf fresh weight. ( f ) Quantification of H 2 O 2 in healthy and Lpsy-infected tomato plants. ( g ) Transmission electron microscopy (TEM) analysis of callose deposition in the midveins of indicated tomato genotypes under healthy and Lpsy-infected conditions. Red arrows indicate sieve pores. SE, sieve element. ( h ) qRT-PCR analysis of SlPUB21 relative expression levels in eds1- edited lines and wild-type tomato under healthy and Lpsy-infected conditions. The actin gene was used as an internal control. Scale bars represent mean ± SD, n=4. Asterisks indicate statistically significant differences by one-way ANOVA and Tukey’s test ( P < 0.01).

Article Snippet: Leaves at the upper parts, without contact with the solution, were detached for imaging in a PerkinElmer IVIS Lumina S5 imager with GFP channel.

Techniques: Real-time Polymerase Chain Reaction, Staining, Infection, Imaging, Fluorescence, Starch, Control, Transmission Assay, Electron Microscopy, Quantitative RT-PCR, Expressing

Journal: Cancers

Article Title: Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

doi: 10.3390/cancers13061351

Figure Lengend Snippet: Primary fibroblast-derived EXOs display an activation status dependent protein cargo. The established fibroblast cell lines were subjected to cellular protein isolation after 48 h incubation of equal cell numbers in normal growth medium. EXOs were isolated using combined differential centrifugation and ultrafiltration approach following 48 h of fibroblast incubation in starvation medium. Exosomal protein was isolated in biological triplicates and subjected to Mass Spectrometry. Statistical Analysis was performed in Perseus. ( A ) Immunoblot analysis of vimentin (VIM), α-smooth-muscle actin (αSMA), fibroblast activation protein α (FAPα), caveolin 1 (CAV1), cluster of differentiation 90 (CD90)/Thy1 and fibroblast-specific protein 1 (FSP1) in primary fibroblasts, including GAPDH as loading control. ( B ) Particle size distribution in the isolated EXOs (mean of n = 4–10) as measured by nanoparticle tracking analysis (NTA) using ZetaView ® . ( C ) Representative transmission electron microscopy (TEM) image of fibroblast-derived EXOs. ( D ) Immunoblot analysis of EXO markers CD9, CD63, CD81, Flotillin 1 and Tumor susceptibility 101 (TSG101), including calreticulin as negative control. ( E ) Heat map of mass spectrometry data illustrating significantly deregulated vesicular proteins. Proteins with more than 4 undefined values in total or more than 3 undefined values in the NF/CAF subgroups are excluded. Paired t test: q < 0.05, diff. > |1.0|. LFQ: label-free quantification; nda: no data acquired. ( F ) Immunoblot analysis of quiescin sulfhydryl oxidase 1 (QSOX1), actinin α4 (ACTN4), thrombospondin 1 (THBS1) and EGF-like repeats and discoidin domains 3 (EDIL3) in EXOs. ( G ) Immunoblot analysis of QSOX1, ACTN4, THBS1 and EDIL3 in primary fibroblasts whole cell lysate, including GAPDH as loading control. The images of uncropped western blot figures are shown in .

Article Snippet: For the detection of proteins of interest, the following primary antibodies were used: recombinant anti-CD9 antibody [EPR2949] (ab92726, Abcam, 1:200), anti-CD63 antibody (ab68418, Abcam, 1:500), CD81 antibody (NBP2-20564, Novus Biologicals, Centennial, CO, USA, 1:400), anti-TSG101 antibody [4A10] (ab83, Abcam, 1:500), Flotillin-1 antibody (3253, Cell Signaling Technology, 1:500), calreticulin antibody (2891, Cell Signaling Technology, 1:1000), recombinant anti-Quiescin Q6 antibody [EPR21866] (ab235444, Abcam, 1:1000), α-actinin 4 (D7U5A) rabbit mAB (15145, Cell Signaling Technology, 1:1000), thrombospondin-1 (D7E5F) rabbit mAb (37879, Cell Signaling Technology, 1:200) and human EDIL3 antibody (MAB6046, R&D Systems, 1:500).

Techniques: Derivative Assay, Activation Assay, Isolation, Incubation, Centrifugation, Mass Spectrometry, Western Blot, Transmission Assay, Electron Microscopy, Negative Control

Journal: Cancers

Article Title: Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

doi: 10.3390/cancers13061351

Figure Lengend Snippet: Selected primary fibroblast activation status dependent EXO markers display specificity to blood EXOs in matched CRC patient plasma. ( A ) Particle size distribution of patient-matched plasma EXOs (pEXO) in comparison to whole plasma (wP) and EXO-depleted plasma (edP) as measured by NTA. ( B ) Immunoblot analysis of quiescin sulfhydryl oxidase 1 (QSOX1), thrombospondin 1 (THBS1), EGF-like repeats and discoidin domains 3 (EDIL3), the EXO markers CD9, CD63 and Flotillin 1, including calreticulin as negative control, and albumin in wP, pEXO and edP protein lysates. ( C ) Graphical analysis of immunoblots shown in (B) using ImageJ, comparing signal strength to pEXO mix. Unpaired t -test: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For the detection of proteins of interest, the following primary antibodies were used: recombinant anti-CD9 antibody [EPR2949] (ab92726, Abcam, 1:200), anti-CD63 antibody (ab68418, Abcam, 1:500), CD81 antibody (NBP2-20564, Novus Biologicals, Centennial, CO, USA, 1:400), anti-TSG101 antibody [4A10] (ab83, Abcam, 1:500), Flotillin-1 antibody (3253, Cell Signaling Technology, 1:500), calreticulin antibody (2891, Cell Signaling Technology, 1:1000), recombinant anti-Quiescin Q6 antibody [EPR21866] (ab235444, Abcam, 1:1000), α-actinin 4 (D7U5A) rabbit mAB (15145, Cell Signaling Technology, 1:1000), thrombospondin-1 (D7E5F) rabbit mAb (37879, Cell Signaling Technology, 1:200) and human EDIL3 antibody (MAB6046, R&D Systems, 1:500).

Techniques: Activation Assay, Western Blot, Negative Control