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