Journal: Communications Biology

Article Title: Exogenous dsRNA triggers sequence-specific RNAi and fungal stress responses to control Magnaporthe oryzae in Brachypodium distachyon

doi: 10.1038/s42003-025-07554-6

Figure Lengend Snippet: Three-day-old Mo liquid cultures were incubated with different RNA molecules for 2 min and subsequently stained for CSLM analysis. A Germinated conidia were treated with 500 ng/µL of poly(I:C) as positive stressor or 10 ng/µL of Pmk1-dsRNA and SHP-siRNA. B Dose-response effect of ROS production in germinating conidia after treatment with SHP-dsRNA. Fluorescence detection with AF488 [λexcitation (nm): 492; λemission (nm): 561]. Scale bar equals 50 µm.

Article Snippet: A silencer siRNA Labeling Kit (ThermoFisher) with Fluorescein dye was used to label Phi6-dsRNA and 21 bp GAPDH siRNA provided with the kit.

Techniques: Incubation, Staining, Fluorescence

Journal: Communications Biology

Article Title: Exogenous dsRNA triggers sequence-specific RNAi and fungal stress responses to control Magnaporthe oryzae in Brachypodium distachyon

doi: 10.1038/s42003-025-07554-6

Figure Lengend Snippet: A Conidia (2000/100 µL) in 0.002% (v/v) Tween20 were incubated with 20 ng/µL of the indicated dsRNA. Potassium chloride (0.25 M) was used as positive stressor control, while geneticin was used as negative control. Images were taken with the AF488 laser at 5 and 60 min after treatment. B Conidia were incubated with 20 ng/µL of the indicated siRNA. C Dose-effect analysis of GFP-MoHog1p nuclear accumulation in Mo conidia treated with various amounts of GFP-dsRNA (476 bp). Scale bar equals 10 µm. AF488 [λexcitation (nm): 492; λemission (nm): 561].

Article Snippet: A silencer siRNA Labeling Kit (ThermoFisher) with Fluorescein dye was used to label Phi6-dsRNA and 21 bp GAPDH siRNA provided with the kit.

Techniques: Incubation, Control, Negative Control

Journal: Communications Biology

Article Title: Exogenous dsRNA triggers sequence-specific RNAi and fungal stress responses to control Magnaporthe oryzae in Brachypodium distachyon

doi: 10.1038/s42003-025-07554-6

Figure Lengend Snippet: ( A ) Infection symptoms on Bd leaves sprayed with a mixture of Mo conidia and dsRNA or siRNA. Intact three-week-old Bd plants were inoculated with a 0.002% (v/v) Tween20 containing conidia (65 × 10³ conidia mL −1 ) and 1 ng/µL of siRNA or long dsRNA. Control plants were sprayed with 0.002% (v/v) Tween (Buffer) or with conidia in Tween solution (Untreated). For imaging, second youngest leaves were detached at 6 dpi and placed on 1% agar plates. Scale bar = 20 mm. B Relative size of necrotic area compared to the whole leaf area calculated with ImageJ. The results of three independent experiments are shown as box plots representing the average with standard deviation. Statistical significance was assessed with Kruskal-Wallis test ( p ≤ 0.05) and asterisks denote difference to the control group (CTR-untreated) according to Dunn’s multiple comparisons test (*: p = 0.046; **: p = 0.0086; ****: p ≤ 0.0001), while differences between GFP and Pmk1 specific RNAs were assessed with two-tailed Welch’s t -test (**: p = 0.0059; ****: p ≤ 0.0001). C Relative fungal growth determined by RT-qPCR comparing Mo housekeeping gene MoGPD with Bd housekeeping gene BdUbi10 . The percentage of reduced fungal growth from three independent experiments was combined and represented as average with standard deviation. Statistical significance assessed with two-tailed t -test (**: p = 0.0013) and one sample t -test to the control (CTR) group (*: p = 0.0207; **: p = 0.0096).

Article Snippet: A silencer siRNA Labeling Kit (ThermoFisher) with Fluorescein dye was used to label Phi6-dsRNA and 21 bp GAPDH siRNA provided with the kit.

Techniques: Infection, Control, Imaging, Standard Deviation, Two Tailed Test, Quantitative RT-PCR

Journal: Communications Biology

Article Title: Exogenous dsRNA triggers sequence-specific RNAi and fungal stress responses to control Magnaporthe oryzae in Brachypodium distachyon

doi: 10.1038/s42003-025-07554-6

Figure Lengend Snippet: A Intact three-week-old Bd seedlings were sprayed with a solution containing conidia (65 × 10³ conidia mL −1 ) and 0.03 ng/µL of the indicated siRNA and dsRNA. Infection symptoms were determined at 6 dpi and the relative size of the necrotic area was quantified using ImageJ. The results of three independent replicates were combined and box plots represent average with standard deviation. Statistical significance was assessed with Kruskal-Wallis test ( p ≤ 0.05) and asterisks denote difference to the control group according to Dunn’s multiple comparisons test (* Pmk1-dsRNA: p = 0.0279; * Pmk1-siRNA: p = 0.0267. B Silencing of the fungal MoPmk1 gene in response to dsRNA treatment of Brachypodium distachyon leaves. Three-week-old Bd seedlings were sprayed with a mixture of Mo conidia and 0.03 ng/µL of fungal target-specific Pmk1-dsRNA, Pmk1-siRNA or GFP -dsRNA or -siRNA. Leaves were harvested at 12 hpi and analysed with RT-qPCR using MoGPD for normalization. Bars represent average of three experiments combined with standard deviation. Statistical significance was assessed with One-way ANOVA test ( p ≤ 0.05) and asterisks denote difference to the control group according to Dunnett test. (** Pmk1-dsRNA: p = 0.0033; ** Pmk1-siRNA: p = 0.0024). Mo indicates a non-treated control.

Article Snippet: A silencer siRNA Labeling Kit (ThermoFisher) with Fluorescein dye was used to label Phi6-dsRNA and 21 bp GAPDH siRNA provided with the kit.

Techniques: Infection, Standard Deviation, Control, Quantitative RT-PCR

Journal: bioRxiv

Article Title: microRNA-210 enhances cell survival and paracrine potential for cardiac cell therapy while targeting mitophagy

doi: 10.1101/2025.01.09.632206

Figure Lengend Snippet: (A) Gapdh mRNA levels following transfection of CPCs with a positive control Gapdh siRNA. (B) Bnip3 mRNA levels following transfection of CPCs with Bnip3 siRNA at 100nM in comparison to a negative siRNA and an untransfected control. Analysed using a one-way ANOVA with Tukey’s multiple comparisons test (n=3).

Article Snippet: The miRNAs and siRNAs used were: mmu-miR-210-3p mirVana miRNA mimic ID: MC10516 (#4464066), mirVana miRNA Mimic Negative Control (#4464059), Silencer Select Pre-Designed siRNA Application Silencer Select Assay ID: S63060 (#4390771), Silencer Select GAPDH Positive Control siRNA (#4390849), Silencer Select Negative Control No. 2 siRNA (#4390846) (ThermoFisher).

Techniques: Transfection, Positive Control, Comparison, Control

Journal: bioRxiv

Article Title: microRNA-210 enhances cell survival and paracrine potential for cardiac cell therapy while targeting mitophagy

doi: 10.1101/2025.01.09.632206

Figure Lengend Snippet: (A) DNA fragmentation represented as TUNEL + cells following serum starvation of CPCs transfected with the following: Bnip3 siRNA+miR-210, Bnip3 siRNA+negative miRNA, negative siRNA+miR-210, negative siRNA+negative miRNA. Cells were imaged using an FV1000 confocal microscope. High power inserts are shown by white boxes. DNase treatment was used as a TUNEL assay positive control. (B) Quantification of the TUNEL assay. (C) mRNA levels of Bnip3 and Nix in miR-210-transfected CPCs, in comparison to CPCs transfected with a negative miRNA, under the following conditions: control media and normoxia, serum starvation and normoxia, control media and hypoxia, serum starvation and hypoxia. Analysed using a one-way ANOVA with Tukey’s multiple comparisons test (n=3); * p < 0.05, ** p < 0.01.

Article Snippet: The miRNAs and siRNAs used were: mmu-miR-210-3p mirVana miRNA mimic ID: MC10516 (#4464066), mirVana miRNA Mimic Negative Control (#4464059), Silencer Select Pre-Designed siRNA Application Silencer Select Assay ID: S63060 (#4390771), Silencer Select GAPDH Positive Control siRNA (#4390849), Silencer Select Negative Control No. 2 siRNA (#4390846) (ThermoFisher).

Techniques: TUNEL Assay, Transfection, Microscopy, Positive Control, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: MicroRNA Profiling of PRELI-Modulated Exosomes and Effects on Hepatic Cancer Stem Cells

doi: 10.3390/ijms252413299

Figure Lengend Snippet: Expression of AKT signaling molecules in LCSCs under miRNA treatment. The merged images show the internalized fluorescence-labeled GAPDH-siRNAs (GFP: green fluorescent proteins) and miRNA (Cy5: cyanine 5) in LCSCs. The expression levels of AKT signaling molecules (phosphorylated AKT and phosphorylated mTORC1) in LCSCs transfected with the fluorescent (Cy5)-labeled miRNAs including hsa-miR378a-3p, hsa-miR25-3p, and hsa-miR423-3p and positive control siRNAs (GFP-GAPDH -siRNA) (* p < 0.05, ** p < 0.01, *** p < 0.001) (scale bars = 20 μm).

Article Snippet: During incubation, LCSCs were transfected with miRNAs using Lipofectamine 2000 reagent (Invitrogen) with control siRNA oligonucleotide (negative) (Bioneer) and GFP-GAPDH siRNA (Bioneer) for one day.

Techniques: Expressing, Fluorescence, Labeling, Transfection, Positive Control