reverse transcription amplification controls  (ATCC)

Journal: medRxiv

Article Title: Bio-Rad and QIAGEN digital PCR platforms provide equivalent quantification for wastewater-based SARS-CoV-2 surveillance

doi: 10.64898/2026.01.20.26344437

Figure Lengend Snippet: Quantitative comparison of SARS-CoV-2 targets between Bio-Rad QX200 ddPCR and QIAGEN QIAcuity dPCR platforms across all concentration ranges. Scatter plots showing hyperwelled (merged triplicate) concentrations for (A) N1 gene target and (B) N2 gene target. Each point represents a single wastewater sample (n = 95) plotted as log 10 copies L -1 on both axes. Samples are color-coded by concentration bin: high (red, 5×10 4 to 5×10 5 copies L -1 , n=31), medium (orange, 5×10 3 to 5×10 4 copies L -1 , n=32), and low (blue, 1×10 3 to 5×10 3 copies L -1 , n=31). Solid black line represents perfect 1:1 agreement; dashed line showed linear regression fit. Linear regression equations, R 2 values, and 95% confidence intervals (gray shading) are displayed for each target. Mean absolute differences between platforms: N1 = 0.10 log copies L -1 , N2 = 0.06 log copies L -1 . All correlations significant at p < 0.001.

Article Snippet: For positive controls, we used genomic SARS-CoV-2 RNA positive control from strain 2019nCoV/USA-WA1/2020 (VR-1986D, ATCC Bethesda, MD), and the positive controls were included with each plate in duplicate for the N1 and N2 assays.

Techniques: Comparison, Concentration Assay

Journal: medRxiv

Article Title: Bio-Rad and QIAGEN digital PCR platforms provide equivalent quantification for wastewater-based SARS-CoV-2 surveillance

doi: 10.64898/2026.01.20.26344437

Figure Lengend Snippet: Platform comparison of SARS-CoV-2 quantification stratified by concentration bin. Box plots showing distribution of log 10 -transformed concentrations (copies L -1 ) for (A) N1 gene target and (B) N2 gene target across low, medium, and high concentration bins. Bio-Rad QX200 data shown in blue; QIAGEN QIAcuity data shown in green. Each box represents the interquartile range (IQR, 25 th -75 th percentile), with the horizontal line indicating the medium. Whiskers extend to 1.5x IQR or the most extreme data point within this range. Individual data points are overlaid as semi-transparent dots to show data distribution (n=31 for high and low bins, n=32 for medium bin). Asterisks indicate statistically significant differences between platforms within each bin as determined by linear mixed effects modeling (*p < 0.05, **p < 0.01, ***p < 0.001). Note that all statistically significant differences are ≤ 0.13 log copies L -1 . Sample numbers are indicated for each concentration bin. Both platforms successfully quantified 100% of samples across all bins.

Article Snippet: For positive controls, we used genomic SARS-CoV-2 RNA positive control from strain 2019nCoV/USA-WA1/2020 (VR-1986D, ATCC Bethesda, MD), and the positive controls were included with each plate in duplicate for the N1 and N2 assays.

Techniques: Comparison, Concentration Assay, Transformation Assay

Journal: medRxiv

Article Title: Bio-Rad and QIAGEN digital PCR platforms provide equivalent quantification for wastewater-based SARS-CoV-2 surveillance

doi: 10.64898/2026.01.20.26344437

Figure Lengend Snippet:

Article Snippet: For positive controls, we used genomic SARS-CoV-2 RNA positive control from strain 2019nCoV/USA-WA1/2020 (VR-1986D, ATCC Bethesda, MD), and the positive controls were included with each plate in duplicate for the N1 and N2 assays.

Techniques:

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: GBD activates Nrf2 signaling in response to LTA. MH-S cells were pretreated with GBD for 30 min and then stimulated with LTA. (A) Confocal images indicating Nrf2 nuclear translocation 3 h poststimulation. Scale bar, 50 µm. (B) Time-course of Nrf2 mRNA expression after LTA stimulation (0, 2, 4, 6, and 8 h). (C) Nrf2 mRNA expression was determined through reverse transcription-quantitative PCR 2 h poststimulation, as described in the materials and methods section. (D) Western blot analysis of Nrf2 protein levels 3 h after LTA stimulation. Data are presented as means±SD. *P<0.05, **P<0.01 and ***P<0.001 vs. 0 h or DMSO; ## P<0.01 and ### P<0.001 vs. DMSO+LTA. GBD glabridin; Nrf2, nuclear factor erythroid 2-related factor 2; LTA, lipoteichoic acid; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Translocation Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: GBD increases HO-1 expression through Nrf2 signaling under LTA challenge. MH-S cells were pretreated with GBD for 30 min and then stimulated with LTA. (A) Confocal images of HO-1 expression 6 h poststimulation. Scale bar, 50 µm. (B) Time-course of HO-1 mRNA expression after LTA stimulation (0, 2, 4, 6, and 8 h). (C) HO-1 mRNA expression was determined by reverse transcription-quantitative PCR. (D) HO-1 protein expression analyzed through western blotting 6 h poststimulation. Data are presented as means±SD (n=4). *P<0.05 and ***P<0.001 vs. 0 h or DMSO; ### P<0.001 vs. DMSO+LTA. GBD glabridin; HO-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2-related factor 2; LTA, lipoteichoic acid; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: ML385 inhibits GBD-induced Nrf2 activation in LTA-stimulated MH-S cells. MH-S cells were pretreated with DMSO (0.1%), GBD (20 µM), or ML385 (5 µM) for 30 min and then stimulated with LTA. (A) Confocal microscopy of Nrf2 nuclear localization 3 h poststimulation. Scale bar, 10 µm. (B) Quantification of Nrf2 nuclear accumulation based on nuclear mean fluorescence intensity. (C) Nrf2 mRNA expression 2 h poststimulation, analyzed through reverse transcription-quantitative PCR. Data are presented as means ± SD. ***P<0.001 vs. DMSO+LTA; ## P<0.01 vs. GBD+LTA; †† P<0.01 and ††† P<0.001 vs. ML385+LTA. GBD glabridin; Nrf2, nuclear factor erythroid 2-related factor 2; LTA, lipoteichoic acid; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Activation Assay, Confocal Microscopy, Fluorescence, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: Pharmacological and genetic inhibition of Nrf2 attenuates GBD-induced HO-1 expression. MH-S cells were pretreated with the indicated compounds for 30 min and then stimulated with LTA. (A) Immunofluorescence analysis of HO-1 expression 6 h poststimulation. Scale bar, 10 µm. (B) Immunofluorescence image evaluation of HO-1 mean fluorescence intensity. (C) HO-1 mRNA expression 6 h poststimulation, determined through RT-qPCR. (D and E) Effect of Nrf2 knockdown on GBD-induced HO-1 expression. MH-S cells were transfected with control siRNA (siNC) or Nrf2 siRNA (siNrf2) for 6 h, followed by the indicated treatments. HO-1 mRNA levels were measured by RT-qPCR. Data are presented as means ± SD. ***P<0.001 vs. DMSO+LTA (B-C) or si-Ctrl+DMSO (D and E); ## P<0.01 and ### P<0.001 vs. GBD+LTA (B-C) or si-Ctrl+DMSO+LTA (E); †† P<0.01 and ††† P<0.001 vs. ML385+LTA (B-C) or si-Ctrl+GBD+LTA (E). Nrf2, nuclear factor erythroid 2-related factor 2; GBD, glabridin; LTA, lipoteichoic acid; HO-1, heme oxygenase-1; RT-qPCR, reverse transcription-quantitative PCR; si, short interfering; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Inhibition, Expressing, Immunofluorescence, Fluorescence, Quantitative RT-PCR, Knockdown, Transfection, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: GBD inhibits LTA-induced macrophage migration through Nrf2 activation. MH-S cells were pretreated with the indicated compounds for 30 min and then stimulated with LTA. (A) Representative wound healing images 0, 6, and 24 h after LTA stimulation. (B) Quantification of wound closure. (C) Representative images of Transwell migration at 6 h and 24 h after LTA stimulation. Migrated cells on the lower surface of the membrane were fixed, stained with crystal violet and images captured under an inverted light microscope. Scale bar, 200 µm. (D) Quantification of migrated cells. Data are presented as means ± SD. ***P<0.001 vs. DMSO; ### P<0.001 vs. DMSO+LTA; † P<0.05, †† P<0.01 and ††† P<0.001 vs. GBD+LTA; ‡‡ P<0.01 vs. ML385+LTA. GBD, glabridin; LTA, lipoteichoic acid; Nrf2, nuclear factor erythroid 2-related factor 2; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Migration, Activation Assay, Membrane, Staining, Light Microscopy

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: Schematic of GBD-induced modulation of macrophage migration in vitro . Upon LTA stimulation, MH-S cells increase ROS production, activating Nrf2 and upregulating HO-1. GBD thus modulateS cell migration by increasing Nrf2 nuclear translocation and HO-1 expression. GBD, glabridin; LTA, lipoteichoic acid; ROS, reactive oxygen species; Nrf2, nuclear factor erythroid 2-related factor 2; HO-1, heme oxygenase-1.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Migration, In Vitro, Translocation Assay, Expressing