hot firepol probe qpcr mix plus (rox)  (Solis BioDyne)

Journal: STAR Protocols

Article Title: Protocol for RNA interference in gene expression in adult insects of Triatoma infestans (Hemiptera: Reduviidae)

doi: 10.1016/j.xpro.2025.103865

Figure Lengend Snippet: Relative expression levels of per mRNA in the heads of adult Triatoma infestans after injection with per-specific double-stranded RNA (dsRNA) Levels of per transcript quantification was performed using quantitative PCR (qPCR), normalizing against β - actin mRNA. Results are displayed as boxplots, with the boxes representing the interquartile range and the whiskers indicating data variability outside the upper and lower quartiles. Asterisks indicate statistically significant differences between the dsRNA per group and controls (∗∗∗p <0.001, ∗∗∗∗p < 0.0001). Control group: group non injected; Control β-Lac dsRNA group: group injected with β-Lac dsRNA; dsRNA per group: group injected with per dsRNA.

Article Snippet: HOT FIREPol Probe qPCR Mix Plus (ROX), 5X , SOLIS BIODYNE , Cat#08-14-00001.

Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction, Control

Journal: Immunity & ageing : I & A

Article Title: Elocalcitol mitigates high-fat diet-induced microglial senescence via miR-146a modulation.

doi: 10.1186/s12979-024-00485-6

Figure Lengend Snippet: Fig. 2 High-fat diet induces senescence in the hypothalamus. (A) Representative dot plots of flow cytometry showing β-gal positive CD45 + CD11b + microglial cells. (B) Quantification of β-gal positive CD45 + CD11b + microglial cells in the hypothalamus. (C-D) RT-qPCR analysis of p16 and p21 mRNA expression. Number of animals = 5–8. Student’s t-test (A, C) and Mann Whitney test (B). Data are presented as mean ± SEM; ** p < 0.01, *** p < 0.001

Article Snippet: For cRNA synthesis, TaqMan® MicroRNA reverse transcription kit (4366596, Thermo Scientific) and for qPCR, 5× HOT FIREPol® Probe qPCR Mix Plus (ROX) (Solis BioDyne) were used, respectively.

Techniques: Flow Cytometry, Quantitative RT-PCR, Expressing, MANN-WHITNEY

Journal: Immunity & ageing : I & A

Article Title: Elocalcitol mitigates high-fat diet-induced microglial senescence via miR-146a modulation.

doi: 10.1186/s12979-024-00485-6

Figure Lengend Snippet: Fig. 3 Microglial activation and elevated SASP cytokines in the hypothalamus induced by HFD. (A) Representative immunohistochemistry microphotographs of the hypothalamic Iba1 + cells. (B) Density of microglial cells. (C) Average cell size of microglial cells. (D) Average cell body size of Iba1 + cells in the hypothalamus. (E) Relative mRNA expression of Il1b (F) Il18 (G) Tnf and (H) Il6 was analyzed by RT-qPCR with primers for indicated genes. Number of animals = 4–8. Student’s t-test (B, C, F, H) and Mann Whitney test (D, E, G). Data represented as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: For cRNA synthesis, TaqMan® MicroRNA reverse transcription kit (4366596, Thermo Scientific) and for qPCR, 5× HOT FIREPol® Probe qPCR Mix Plus (ROX) (Solis BioDyne) were used, respectively.

Techniques: Activation Assay, Immunohistochemistry, Expressing, Quantitative RT-PCR, MANN-WHITNEY

Journal: Immunity & ageing : I & A

Article Title: Elocalcitol mitigates high-fat diet-induced microglial senescence via miR-146a modulation.

doi: 10.1186/s12979-024-00485-6

Figure Lengend Snippet: Fig. 4 High-fat diet induces senescence-associated changes and SASP cytokines in the liver. (A-B) Relative mRNA expression and protein levels of p16, (C-D) Relative mRNA expression and protein levels of p21 and (E-G) Relative mRNA expression of Il18, Tnf, Il6 cytokines, analyzed by RT-qPCR. Number of animals = 5. Student’s t-test (A-E, G) and Mann Whitney test (F). Data represented as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: For cRNA synthesis, TaqMan® MicroRNA reverse transcription kit (4366596, Thermo Scientific) and for qPCR, 5× HOT FIREPol® Probe qPCR Mix Plus (ROX) (Solis BioDyne) were used, respectively.

Techniques: Expressing, Quantitative RT-PCR, MANN-WHITNEY

Journal: Immunity & ageing : I & A

Article Title: Elocalcitol mitigates high-fat diet-induced microglial senescence via miR-146a modulation.

doi: 10.1186/s12979-024-00485-6

Figure Lengend Snippet: Fig. 7 HFD induces senescence markers and SASP cytokines in the liver of WT and Mir146a-/- mice. qPCR analysis of (A) p16 mRNA expression, (B) p21 mRNA expression and (C-E) RT-qPCR analysis of SASP cytokines: Il18, Tnf, and Il6 in the liver of WT and Mir146a-/- mice on HFD and LFD. Number of animals = 5. Data panels (A, C, D, E) were analyzed using two-way ANOVA with Tukey’s multiple comparisons test, data in panel (B) did not pass normality test and transformed into logarithms (Y = log(Y) and further processed using one-way ANOVA with Tukey’s multiple comparisons test. Data are represented as mean ± SEM; * p < 0.05, ** p < 0.01

Article Snippet: For cRNA synthesis, TaqMan® MicroRNA reverse transcription kit (4366596, Thermo Scientific) and for qPCR, 5× HOT FIREPol® Probe qPCR Mix Plus (ROX) (Solis BioDyne) were used, respectively.

Techniques: Expressing, Quantitative RT-PCR, Transformation Assay

Journal: Immunity & ageing : I & A

Article Title: Elocalcitol mitigates high-fat diet-induced microglial senescence via miR-146a modulation.

doi: 10.1186/s12979-024-00485-6

Figure Lengend Snippet: Fig. 8 Smad4 mRNA expression is increased in the hypothalamus of Mir146a-/- mice compared to WT mice in HFD conditions. (A, B) RT-qPCR analysis of Irak1 and Smad4 mRNA expression levels in the hypothalamus and (C, D) liver. Number of animals = 4–6. All data panels were analyzed using two-way ANOVA followed by Tukey’s multiple comparisons test. Data are represented as mean ± SEM; * p < 0.05, ** p < 0.01

Article Snippet: For cRNA synthesis, TaqMan® MicroRNA reverse transcription kit (4366596, Thermo Scientific) and for qPCR, 5× HOT FIREPol® Probe qPCR Mix Plus (ROX) (Solis BioDyne) were used, respectively.

Techniques: Expressing, Quantitative RT-PCR

Journal: Immunity & ageing : I & A

Article Title: Elocalcitol mitigates high-fat diet-induced microglial senescence via miR-146a modulation.

doi: 10.1186/s12979-024-00485-6

Figure Lengend Snippet: Fig. 10 Elocalcitol reduces HFD-induced senescence and SASP cytokines in liver of WT mice but not Mir146a-/- mice. (A) qPCR analysis of p21 mRNA expression levels, (B) Relative mRNA expression of Il18, (C) Tnf, and (D) Il6 in the liver, analyzed by RT-qPCR with primers for the indicated genes in the liver. Number of animals = 4–5. Data in panels (A-C) did not show normal distribution and were transformed into logarithms (Y = log(Y) and further processed using one-way ANOVA with Tukey’s multiple comparisons test and (D) was analyzed using two-way ANOVA followed by Tukey’s multiple comparisons test. Data are represented as mean ± SEM; * p < 0.05, ** p < 0.01

Article Snippet: For cRNA synthesis, TaqMan® MicroRNA reverse transcription kit (4366596, Thermo Scientific) and for qPCR, 5× HOT FIREPol® Probe qPCR Mix Plus (ROX) (Solis BioDyne) were used, respectively.

Techniques: Expressing, Quantitative RT-PCR, Transformation Assay