Journal: Frontiers in Microbiology

Article Title: Intestinal Flora-Derived Kynurenic Acid Protects Against Intestinal Damage Caused by Candida albicans Infection via Activation of Aryl Hydrocarbon Receptor

doi: 10.3389/fmicb.2022.934786

Figure Lengend Snippet: KynA suppressed the MK2/p-MK2 signaling pathway by activating AHR. We investigated the regulation of TTP and expression changes of MK2 and P-MK2 induced by AHR activation at the cellular level. In Caco-2 cells, after treatment with KynA, expression of CYP1A1 increased significantly, as did expression of TTP also ( P < 0.05), while expression of MK2 decreased significantly ( P < 0.05). In the CA- Caco-2 cell model, the expression of proteins CYP1A1 and TTP decreased significantly, but the expression of p-MK2 increased ( P < 0.01). After the treatment of Caco-2 cells with KynA and the AHR agonist FICZ, the expression of CYP1A1 and TTP was significantly increased, but the expression of p-MK2 decreased significantly ( P < 0.01). Treatment with an AHR inhibitor (CH223191) led to a decrease in the expression of CYP1A1 and TTP, and an increase in the expression of p-MK2 ( P < 0.01). Statistical significance was evaluated using the Mann–Whitney U test. P -values < 0.05 (*) or < 0.01 (**) were considered statistically significant. CA, Candida albicans infection; KynA, Kynurenic acid; FICZ, 6-Formylindolo[3,2-b]carbazole. P -values < 0.001 (***).

Article Snippet: Proteintech, United States); anti-AHR antibody (1:1,000, 67785-1-Ig; Proteintech, United States); anti-ZO-1 antibody (1:1,000, 21773-1-AP, Proteintech, United States); anti-GAPDH antibody (1:1,000, 60004-1-Ig, Proteintech, United States); anti-MK2 antibody (1:1,000, 13949-1-AP, Proteintech, United States); anti-CYP1A1 antibody (1:1,000, 13241-1-AP, Proteintech, United States); anti-TTP antibody (1:1,000, 12737-1-AP; Proteintech, United States); anti-MLCK antibody (1:1,000, 21642-1-AP; Proteintech, United States); and anti-pMLC antibody (1:2,000, CST-3671; Cell Signaling Technology, United States).

Techniques: Expressing, Activation Assay, MANN-WHITNEY, Infection

Journal: Nucleic Acids Research

Article Title: Ectopic over-expression of tristetraprolin in human cancer cells promotes biogenesis of let-7 by down-regulation of Lin28

doi: 10.1093/nar/gkr1302

Figure Lengend Snippet: TTP negatively regulates the levels of Lin28a in human cancer cells. ( A–D ) Overexpression of TTP inhibits Lin28a levels in PA1 cells. PA1 cells were transfected with pcDNA6/V5-TTP or pcDNA6/V5 for 24 h. (A) The levels of TTP, Drosha, Dicer, Ago2 and Lin28a proteins were determined by western blot assays. (B) The expression levels of TTP and Lin28a were determined by semi-qRT-PCR. (C) The level of Lin28a was determined by qRT-PCR. The levels obtained from PA1/pcDNA cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (** P < 0.01). (D) Overexpression of TTP decreases Lin28b levels in PA1 cells. The level of TTP and Lin28b were determined by RT-PCR (top panel) and western blot (bottom panel). ( E–I ) Downregulation of TTP by siRNA increases Lin28a levels and decreases let-7b in HCT116 cells. HCT116 cells were transfected with TTP-specific (TTP-siRNA) or scRNA. After 24 h, the levels of TTP and Lin28a were determined by qRT-PCR (E and F) and western blot assays (G). The level of let-7b was determined by qRT-PCR (H) and cell viability was assessed by measuring absorbance at 490 nm using a MTS cell proliferation assay (I). The levels obtained from mock-transfected HCT116 cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (** P < 0.01; *** P < 0.001). ns, not significant. ( J ) The level of TTP protein is inversely correlated with those of Lin28a protein within several human cell lines. Levels of TTP and Lin28a proteins were determined by western blot analysis in PA1 (ovarian teratocarcinoma), MCF7 (breast adenocarcinoma), AGS (gastric adenocarcinoma), K562 (erythroleukemia), HepG2 (hepatocellular carcinoma) and NT2 (neuronally committed teratocarcinoma) cells. β-Actin was detected as the loading control. ( K ) TTP expression level is inversely correlated with that of Lin28a in human ovarian tissues. Representative TTP and Lin28a immunohistochemical staining in normal and ovarian adenocarcinoma tissues. Normal ovarian tissues showed strong immunoreactivity for TTP, where as the ovarian adenocarcinoma showed strong positive staining for Lin28a.

Article Snippet: After deparaffinization, primary antibody anti-human TTP antibody (sc-14030, Santa Cruz Biotechnology) or anti-human Lin28a antibody (ab46020, Abcam) at a 1:100 dilution was applied for 2°h at room temperature.

Techniques: Over Expression, Transfection, Western Blot, Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Proliferation Assay, Control, Immunohistochemical staining, Staining

Journal: Nucleic Acids Research

Article Title: Ectopic over-expression of tristetraprolin in human cancer cells promotes biogenesis of let-7 by down-regulation of Lin28

doi: 10.1093/nar/gkr1302

Figure Lengend Snippet: TTP and Lin28a expression and clinicopathologic features of patients with ovarian adenocarcinoma

Article Snippet: After deparaffinization, primary antibody anti-human TTP antibody (sc-14030, Santa Cruz Biotechnology) or anti-human Lin28a antibody (ab46020, Abcam) at a 1:100 dilution was applied for 2°h at room temperature.

Techniques: Expressing, Significance Assay, Staining

Journal: Nucleic Acids Research

Article Title: Ectopic over-expression of tristetraprolin in human cancer cells promotes biogenesis of let-7 by down-regulation of Lin28

doi: 10.1093/nar/gkr1302

Figure Lengend Snippet: TTP enhances the decay of Lin28a mRNA through interaction with an ARE within the Lin28a mRNA 3′-UTR. ( A and B ) TTP destabilizes Lin28a mRNA. PA1 cells were transfected with pcDNA6/V5-TTP or pcDNA6/V5 for 24 h. (A) The level of TTP protein was determined by western blot assays. (B) Expression of Lin28a mRNA in PA1 cells was determined by qRT-PCR and mRNA half-life was calculated from the non-linear regression of the mRNA levels at the indicated times after the addition of 5.0 µg/ml actinomycin D. A one-phase model of exponential decay was used to derive the indicated mRNA decay curves. Results shown on the graph represent the means ± SD of three independent experiments (** P < 0.01; *** P < 0.001). ( C ) Inhibition of TTP by siRNA enhances Lin28a mRNA stability. HCT116 cells were transfected with siRNA against TTP-specific (TTP-siRNA) or scRNA for 24 h. Expression of Lin28a mRNA in HCT116 cells was determined by qRT-PCR and mRNA half-life was calculated as described in (B). Results shown on the graph represent the means ± SD of three independent experiments (* P < 0.05). ( D ) Lin28a mRNA half-life in PA1 cell with low TTP level is longer than that in AGS cells with high TTP level. Expression of Lin28a mRNA in PA1 and AGS cells was determined by qRT-PCR and mRNA half-life was calculated as described in (B). Results shown on the graph represent the means ± SD of three independent experiments (*** P < 0.001). ( E and F ) The first AUUUA pentamer (ARE1) within the Lin28a 3′-UTR is essential for the inhibitory effect of TTP. (E) Schematic representation of the luciferase reporter constructs used in this study. Fragments (Frag) and oligonucleotides (Oligo) derived from the Lin28a mRNA 3′-UTR were cloned downstream of the luciferase reporter gene in the psiCHECK2 luciferase expression vector. White circles, wild-type (W) pentameric motif AUUUA; gray circles, mutated (M) motif AGCA. (F) PA1 cells were co-transfected with pcDNA6/V5-TTP and a psiCHECK2 luciferase reporter constructs containing fragments (left panel) or oligonucleotides (right panel) derived from the Lin28a mRNA 3′-UTR as described in (E) for 24 h. After normalizing luciferase activity, the luciferase activity obtained from PA1 cells transfected with the Frag-ARE1 luciferase construct alone or Oligo-ARE1W alone were set to 1.0. Results shown represent the means ± SD of three independent experiments (* P < 0.05; ** P < 0.01). ns, not significant. ( G and H ) TTP decreases the luciferase activity of luciferase reporter gene containing the Lin28b 3′-UTR. (G) Schematic representation of the luciferase reporter constructs used in this study. Fragments (Frag) derived from the Lin28b mRNA 3′-UTR were cloned downstream of the luciferase reporter gene in the psiCHECK2 luciferase expression vector. White circles, wild-type pentameric motif AUUUA. (H) PA1 cells were co-transfected with pcDNA6/V5-TTP and a psiCHECK2 luciferase reporter constructs containing fragments derived from the Lin28b mRNA 3′-UTR as described in (G) for 24 h. After normalizing luciferase activity, the luciferase activity obtained from PA1 cells transfected with each Frag-ARE luciferase construct alone was set to 1.0. Results shown represent the means ± SD of three independent experiments (** P < 0.01; *** P < 0.001). ( I ) Ribonucleoprotein immunoprecipitation assay. PA1 cells were cotransfected with pcDNA6/V5-TTP and psiCHECK2 luciferase reporter constructs containing Lin28a Oligo-ARE1W. psiCHECK2 luciferase reporter construct containing mutant ARE1, Oligo-ARE1M was used as a negative control. At 24 h after transfection, the ribonucleoprotein complexes containing TTP were immunoprecipitated with protein G-agarose and anti-V5 or a control antibody. The luciferase mRNA in the immunoprecipitates was amplified by RT-PCR. The presence of TTP in the immunoprecipitates was detected by western blot with anti-V5 antibody. ( J and K ) RNA EMSA was performed by mixing cytoplasmic extracts containing 3.0 µg of total protein from pcDNA6/V5-TTP-transfected PA1 cells (J) or HCT116 cells (K) with 80 fmol biotinylated wild-type Oligo-ARE1W (WT) or mutant Oligo-ARE1M (MUT) probe. Anti-V5 (J), anti-TTP (K) or control antibody was added to the reaction mixtures. Position of the TTP containing bands (TTP) and super-shifted bands (SS) are indicated.

Article Snippet: After deparaffinization, primary antibody anti-human TTP antibody (sc-14030, Santa Cruz Biotechnology) or anti-human Lin28a antibody (ab46020, Abcam) at a 1:100 dilution was applied for 2°h at room temperature.

Techniques: Transfection, Western Blot, Expressing, Quantitative RT-PCR, Inhibition, Luciferase, Construct, Derivative Assay, Clone Assay, Plasmid Preparation, Activity Assay, Immunoprecipitation, Mutagenesis, Negative Control, Control, Amplification, Reverse Transcription Polymerase Chain Reaction

Journal: Nucleic Acids Research

Article Title: Ectopic over-expression of tristetraprolin in human cancer cells promotes biogenesis of let-7 by down-regulation of Lin28

doi: 10.1093/nar/gkr1302

Figure Lengend Snippet: Overexpression of Lin28a attenuates the effects of TTP on let-7 b levels, CDC34 levels and PA1 cell growth. ( A–C ) Downregulation of Lin28a by siRNA increases let-7b levels but decreases CDC34 levels in PA1 cells. PA1 cells were transfected with Lin28a -specific (Lin28a-siRNA) or scRNA for 24 h. (A) The level of Lin28a was determined by qRT-PCR (top panel) and western blot assays (bottom panel). The levels obtained from PA1/scRNA cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (*** P < 0.001). (B) The level of mature let-7b was measured by qRT-PCR. The levels obtained from PA1/scRNA cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (* P < 0.05). (C) The level of CDC34 was determined by qRT-PCR. The levels obtained from PA1/scRNA cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (*** P < 0.001). ( D–F ) Transfection of Lin28a cDNA without the 3′-UTR abolishes the effects of TTP on expression of let-7b , CDC34 and PA1 cell growth. PA1 cells were transfected with a combination of pcDNA6/V5-TTP and pcDNA3/Flag-Lin28a for 24 h. (D) The levels of TTP and Lin28a were measured by semi-qRT-PCR (top panel) and western blot assays (bottom panel). (E) The level of let-7b was measured by qRT-PCR. The levels obtained from PA1/pcDNA cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (* P < 0.05; ** P < 0.01). (F) Cell viability was assessed by measuring absorbance at 490 nm using a MTS cell proliferation assay. The levels obtained from PA1/pcDNA cells were set to 1.0. Data are presented as the mean ± SD ( n = 3) (** P < 0.01; *** P < 0.001).

Article Snippet: After deparaffinization, primary antibody anti-human TTP antibody (sc-14030, Santa Cruz Biotechnology) or anti-human Lin28a antibody (ab46020, Abcam) at a 1:100 dilution was applied for 2°h at room temperature.

Techniques: Over Expression, Transfection, Quantitative RT-PCR, Western Blot, Expressing, Proliferation Assay

Journal: Oncotarget

Article Title: Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells

doi: 10.18632/oncotarget.16706

Figure Lengend Snippet: ( A – E ) SHSY5Y and/or HeLa cells were transiently transfected with pcDNA6/V5-TTP (SHSY5Y/TTP and HeLa/TTP) or with empty vector pcDNA6/V5 (SHSY5Y/pcDNA and HeLa/pcDNA) for 48 h. (A) TTP levels were determined by RT-PCR (top) and western blot (bottom). (B and C) Cells were stained with Mitotracker CMXRos for 30 min, and then images were obtained by confocal microscopy. (B) Representative confocal images with magnified insets of boxed areas. Scale bar, 10 μm. (C) Graphs represent percentage of cells with elongated mitochondria. Values are mean ± s.e.m. from three separate experiments with 100 cells per group per experiment (*** p < 0.001). (D) Representative electron microscopic images of mitochondria. Scale bar, 1 μm. (E) Graphs represent ratio of maximum axis to minimum axis of mitochondria. Values are mean ± s.e.m. from three separate experiments (** p < 0.01). ( F – I ) TTP inhibition induced mitochondrial fragmentation. SHSY5Y and HeLa cells were transiently transfected with scRNA (SHSY5Y/scRNA and HeLa/scRNA) or TTP-siRNA (SHSY5Y/TTP-siRNA and HeLa/TTP-siRNA) for 48 h. (F) TTP levels were determined by RT-PCR (top) and western blot (bottom). The band densities in the western blot were quantified by Image J, normalized to the internal control β-actin and expressed as ratio of the value of control cells. Data shown are mean ± s.e.m. ( n = 3). (*** p < 0.001). (G–I) Cells were stained with Mitotracker CMXRos for 30 min, and images were obtained by confocal microscopy. (G) Representative confocal images with magnified insets of boxed areas. Scale bar, 10 μm. (H) Graphs represent percentage of cells with (top) elongated and (bottom) fragmented mitochondria. Values are mean ± s.e.m. from three separate experiments with 100 cells per group per experiment (* p < 0.05; ** p < 0.01; (*** p < 0.001). (I) Graphs represent ratio of maximum axis to minimum axis of mitochondria. Values are mean ± s.e.m. from three separate experiments (* p < 0.05).

Article Snippet: Small interfering RNAs (siRNAs) against human α-Syn ( α-Syn -siRNA, sc-29619), human TTP ( TTP -siRNA, sc-36760) and control siRNA (scRNA, sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Transfection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Confocal Microscopy, Inhibition, Control

Journal: Oncotarget

Article Title: Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells

doi: 10.18632/oncotarget.16706

Figure Lengend Snippet: ( A ) TTP overexpression did not decrease the expression levels of Mfn1, Mfn2, OPA1, Drp1, and Fis1. SHSY5Y and HeLa cells were transiently transfected with pcDNA6/V5-TTP (SHSY5Y/TTP and HeLa/TTP) or empty vector pcDNA6/V5 (SHSY5Y/pcDNA and HeLa/pcDNA) for 48 h. Cells were analyzed for protein levels by western blot. ( B and C ) TTP overexpression decreased α-Syn levels. SHSY5Y and HeLa cells were transiently transfected with pcDNA6/V5-TTP (SHSY5Y/TTP and HeLa/TTP) or empty vector pcDNA6/V5 (SHSY5Y/pcDNA and HeLa/pcDNA) for 48 h. The levels of TTP and α-Syn were determined by RT-PCR (B, top), western blotting (B, bottom), and qRT-PCR (C). Values are mean ± s.e.m. from three separate experiments (*** p < 0.001). ( D and E ) Inhibition of TTP increased α-Syn levels. SHSY5Y and HeLa cells were transiently transfected with scRNA (SHSY5Y/scRNA and HeLa/scRNA) or TTP-siRNA (SHSY5Y/TTP-siRNA and HeLa/TTP-siRNA) for 48 h. The levels of TTP and α-Syn were determined by RT-PCR (D, top) and western blot (D, bottom). (E) The band densities in the western blot were quantified by Image J, normalized to the internal control β-actin and expressed as ratio of the value of control cells. Values are mean ± s.e.m. from three separate experiments (*** p < 0.001).

Article Snippet: Small interfering RNAs (siRNAs) against human α-Syn ( α-Syn -siRNA, sc-29619), human TTP ( TTP -siRNA, sc-36760) and control siRNA (scRNA, sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Over Expression, Expressing, Transfection, Plasmid Preparation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Inhibition, Control

Journal: Oncotarget

Article Title: Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells

doi: 10.18632/oncotarget.16706

Figure Lengend Snippet: ( A – C ) Inhibition of α-Syn induced elongation of mitochondria. SHSY5Y and HeLa cells were transiently transfected with scRNA (SHSY5Y/scRNA and HeLa/scRNA) or α-Syn-siRNA (SHSY5Y/α-Syn-siRNA and HeLa/α-Syn-siRNA) for 48 h. (A) The levels of α-Syn determined by RT-PCR (top) and western blot (bottom). (B and C) Cells were stained with Mitotracker CMXRos for 30 min, and images were obtained by confocal microscopy. (B) Representative confocal images with magnified insets of boxed areas. Scale bar, 10 μm. (C) Graphs represent percentage of cells with elongated mitochondria. Values are mean ± s.e.m. from three separate experiments with 100 cells per group per experiment (*** p < 0.001). ( D – G ) Transfection of α-Syn cDNA without the 3′-UTR abolished the effects of TTP on mitochondrial morphology. SHSY5Y and/or HeLa cells transfected with a combination of pcDNA6/V5-TTP and pcDNA/α-Syn for 48 h. Cells were stained with Mitotracker CMXRos for 30 min, and images were obtained by confocal microscopy. (D) The levels of TTP and α-Syn were measured by RT-PCR (top panel) and western blot assays (bottom panel). (E) Representative confocal images with magnified insets of boxed areas. Scale bar, 10 μm. (F) Graphs represent percentage of cells with elongated mitochondria. Values are mean ± s.e.m. from three separate experiments with 100 cells per group per experiment (*** p < 0.001). (G) Representative electron microscopic images of mitochondria. Scale bar, 1 μm. (E) Graphs represent ratio of maximum axis to minimum axis of mitochondria. Values are mean ± s.e.m. from three separate experiments (* p < 0.05; ** p < 0.01).

Article Snippet: Small interfering RNAs (siRNAs) against human α-Syn ( α-Syn -siRNA, sc-29619), human TTP ( TTP -siRNA, sc-36760) and control siRNA (scRNA, sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Inhibition, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Confocal Microscopy

Journal: Oncotarget

Article Title: Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells

doi: 10.18632/oncotarget.16706

Figure Lengend Snippet: ( A ) Schematic representation of the luciferase reporter constructs used in this study. ( B ) TTP destabilized α-Syn mRNAs. 2 × 10 6 SHSY5Y cells were transfected with 6 μg pcDNA6/V5-TTP or pcDNA6/V5 for 24 h. The expression of α-Syn mRNAs in SHSY5Y cells was determined by qRT-PCR at the indicated times after addition of 5 μg/ml actinomycin D. Data are presented as the mean ± s.e.m. ( n = 3) (*** p < 0.001). ( C – E ) The second AUUUA pentamer (ARE-2) within the α-Syn 3′-UTR is necessary for the inhibitory effect of TTP. Fragments and oligonucleotides (Oligo) derived from the α-Syn mRNA 3′-UTR were cloned downstream of the luciferase reporter gene in the psiCHECK2 luciferase expression vector. White circles, wild-type (WT) pentameric motif AUUUA; black circles, mutated (MUT) motif AGCA. SHSY5Y cells were co-transfected with pcDNA6/V5-TTP and psiCHECK2 luciferase reporter constructs containing fragments or oligonucleotides derived from the α-Syn mRNA 3′-UTR as described in (A) for 24 h. After normalizing for luciferase activity, the luciferase activity obtained from the SHSY5Y cells transfected with the psiCHECK2 vector alone were set to 1.0. Data are presented as the mean ± s.e.m. ( n = 3) (** p < 0.01; *** p < 0.001). ( F ) Ribonucleoprotein immunoprecipitation assay. SHSY5Y cells were cotransfected with pcDNA6/V5-TTP and psiCHECK2 luciferase reporter constructs containing α-Syn Oligo-ARE-2W. The psiCHECK2 luciferase reporter construct containing mutant ARE-2, Oligo-ARE-2M was used as a negative control. At 48 h after transfection, the ribonucleoprotein complexes containing TTP were immunoprecipitated with protein G-agarose and anti-V5 or a control antibody. Luciferase mRNA in the immunoprecipitates was amplified by RT-PCR. The presence of TTP in the immunoprecipitates was detected by western blot with anti-V5 antibody. ( G ) RNA EMSA was performed by mixing cytoplasmic extracts containing 5 μg of total protein from pcDNA6/V5-TTP-transfected SHSY5Y cells with 20 fmol biotinylated wild-type Oligo-ARE-2W (WT) or mutant Oligo-ARE-2M (MUT) probe. Anti-TTP or control antibody was added to the reaction mixtures. The positions of the TTP-containing bands (TTP) are indicated.

Article Snippet: Small interfering RNAs (siRNAs) against human α-Syn ( α-Syn -siRNA, sc-29619), human TTP ( TTP -siRNA, sc-36760) and control siRNA (scRNA, sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Luciferase, Construct, Transfection, Expressing, Quantitative RT-PCR, Derivative Assay, Clone Assay, Plasmid Preparation, Activity Assay, Immunoprecipitation, Mutagenesis, Negative Control, Control, Amplification, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Oncotarget

Article Title: Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells

doi: 10.18632/oncotarget.16706

Figure Lengend Snippet: ( A and B ) Effects of TTP overexpression on mitochondrial membrane potential. SHSY5Y and HeLa cells were transfected with (A) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (B) scRNA or α-Syn-siRNA for 48 h. Cells were stained with TMRM (pseudo-green) and Mitotracker CMXRos (red) for 30 min, and images were obtained by confocal microscopy. TMRM intensity reflects mitochondrial membrane potential level. The panels display representative confocal images with magnified insets of boxed areas. Scale bar, 10 μm. Graphs represent relative TMRM intensity. Values are mean ± s.e.m. from three separate experiments with 15 cells per group per experiment (** p < 0.01; *** p < 0.001). ( C and D ) Effects of TTP overexpression on ATP production. Cellular ATP levels were measured in SHSY5Y and HeLa cells transfected with (C) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (D) scRNA or α-Syn -siRNA using the CellTiter-Glo luminescent cell viability assay. Values are means ± s.e.m. from three independent experiments (* p < 0.05; ** p < 0.01; *** p < 0.001). ( E and F ) Effects of TTP overexpression on ROS production. ROS production was measured in SHSY5Y and HeLa cells transfected with (E) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (F) scRNA or α-Syn -siRNA using the ROS-Glo ™ H 2 O 2 Assay Kit. Values are means ± s.e.m. of three independent experiments (* p < 0.05; ** p < 0.01; *** p < 0.001). ( G and H ) Effects of TTP overexpression on mtDNA. SHSY5Y and HeLa cells were transfected with (G) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (H) scRNA or α-SYN -siRNA for 48 h. Cells were stained with anti-DNA antibody for mtDNA (green) and DAPI for nuclear DNA (blue) as described in Materials and Methods. The panels display representative confocal images. Graphs represent relative percentage of mtDNA stain remaining. Values are mean ± s.e.m. from three separate experiments with 100 cells per group per experiment (* p < 0.05; ** p < 0.01; *** p < 0.001). Scale bar, 10 μm. ( I and J ) TTP overexpression inhibits oxygen consumption rate (OCR). HeLa cells were transfected with a combination of pcDNA6/V5-TTP and pcDNA/α-Syn for 48 h. OCR of cells were measured by the XF-96 Extracellular Flux Analyzer. OCR was measured under basal conditions followed by the sequential addition of oligomycin (1 μM), FCCP (1 μM) or antimycin A (1 μM). Each data point represents an OCR measurement. (I) Representative traces of OCR measurements. (J) Graphs represent basal respiration, proton leak, and ATP production. Values are mean ± s.e.m. from three separate experiments (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Small interfering RNAs (siRNAs) against human α-Syn ( α-Syn -siRNA, sc-29619), human TTP ( TTP -siRNA, sc-36760) and control siRNA (scRNA, sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Over Expression, Membrane, Transfection, Staining, Confocal Microscopy, Cell Viability Assay

Journal: Oncotarget

Article Title: Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells

doi: 10.18632/oncotarget.16706

Figure Lengend Snippet: ( A and B ) Effects of TTP overexpression on cytochrome release. HeLa cells were transfected with a combination of pcDNA6/V5-TTP and pcDNA/α-Syn for 48 h. Cells were stained with Mitotracker CMXRos (red) and anti-cytochrome c (green), then images were obtained by confocal microscopy. (A) Representative confocal images with magnified insets of boxed areas. Scale bar, 10 μm. (B) The cytosolic fraction of cells was analyzed for cytochrome c by western blot analysis. Graphs represent relative percentage of cytosolic cytochrome c. Values are mean ± s.e.m. from three separate experiments (* p < 0.05; ** p < 0.01). ( C and D ) Effects of TTP overexpression on caspase-3 activity. HeLa cells were transfected with (C) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (D) scRNA or α-Syn -siRNA for 48 h. Cells were analyzed for active caspase-3 using the Image-iT LIVE Red Caspase Detection Kit. Blue, DAPI. Scale bar, 10 μm. Graphs represent percentage of caspase-3-positive cells. Values are mean ± s.e.m. from three separate experiments with 100 cells per group per experiment (** p < 0.01). ( E and F ) Effects of TTP overexpression on apoptosis. HeLa cells were transfected with (E) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (F) scRNA or α-Syn -siRNA for 48 h. Apoptotic cells were analyzed with Annexin V staining by flow cytometric analysis. The panels display representative histogram (** p < 0.01). ( G and H ) Effects of TTP overexpression on cell proliferation. SHSY5Y and HeLa cells were transfected with (G) a combination of pcDNA6/V5-TTP and pcDNA/α-Syn or (H) scRNA or α-Syn -siRNA for 48 h. Cells were seeded in triplicate in 24-well culture plates at 5 × 10 4 cells/well and incubated for 24 h. Cells were then stained by Trypan-Blue Solution (0.4%) (T8154, Sigma) and counted using a hematocytometer. Graphs represent relative cell proliferation. Values are mean ± s.e.m. from three separate experiments (* p < 0.05; *** p < 0.001).

Article Snippet: Small interfering RNAs (siRNAs) against human α-Syn ( α-Syn -siRNA, sc-29619), human TTP ( TTP -siRNA, sc-36760) and control siRNA (scRNA, sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Over Expression, Transfection, Staining, Confocal Microscopy, Western Blot, Activity Assay, Incubation

Journal: Autophagy

Article Title: RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells.

doi: 10.1080/15548627.2019.1687985

Figure Lengend Snippet: Figure 1. RNA-binding protein ZFP36 expression is decreased during HSC ferroptosis. HSC-LX2 and HSC-T6 cells were treated with erastin (10 μM), sorafenib (10 μM), and RSL3 (2.5 μM), with or without the indicated inhibitors (liproxstatin-1, 100 nM; ZVAD-FMK, 10 μM; necrostatin-1, 10 μM) for 24 h. (A) Cell viability, MDA, and iron levels were assayed (n = 3 in every group, **, p < 0.01). (B and C) HSC-LX2 and HSC-T6 cells were treated with erastin (10 μM), sorafenib (10 μM), and RSL3 (2.5 μM) for 24 h. ZFP36 protein and mRNA levels were determined (n = 3 in every group, *, p < 0.05, **, p < 0.01, ***, p < 0.001). (D) HSC-LX2 and HSC-T6 cells were treated with sorafenib (10 μM) with or without MG-132 (10 μM) for 24 h. Cells were harvested for ubiquitination assay (n = 3 in every group). (E) HSC-LX2 cells were treated with erastin (10 μM) with or without cycloheximide (CHX, 20 μg/ml) or MG-132 (10 μM) for 24 h, and ZFP36 protein level was assayed (n = 3 in every group, *, p < 0.05, **, p < 0.01). (F) HSC-LX2 cells were treated with CHX (20 μg/ml) for 24 h or treated with erastin (10 μM) and CHX (20 μg/ml) for 24 h. ZFP36 protein levels were determined at the indicated time points (n = 3 in every group).

Article Snippet: ZFP36 shRNA plasmid (sc-36760-SH, sc-36761-SH), FBXW7 shRNA plasmid (sc-37547-SH, sc-37548-SH) and empty vector were purchased from Santa Cruz Biotechnology.

Techniques: RNA Binding Assay, Expressing, Ubiquitin Proteomics

Journal: Autophagy

Article Title: RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells.

doi: 10.1080/15548627.2019.1687985

Figure Lengend Snippet: Figure 2. The ubiquitin ligase FBXW7 decreases ZFP36 protein expression by recognizing SFSGLPS motif. (A) Amino acid sequence alignment between human and rat ZFP36 protein was determined. (B) HSC-LX2 cells were treated with erastin (10 μM), sorafenib (10 μM), and RSL3 (2.5 μM) for 24 h. The binding of ZFP36 and FBXW7 was determined by immunoprecipitation assay (n = 3 in every group). (C and D) FBXW7-deficient HSC-LX2 and HSC-T6 cells were treated with erastin (10 μM) for 24 h. The ubiquitylation of ZFP36 and the protein levels of ZFP36 and FBXW7 were determined (n = 3 in every group). (E) FBXW7-deficient HSC-LX2 cells were treated with erastin (10 μM) for 24 h. the mRNA levels of ZFP36 and FBXW7 were determined (n = 3 in every group, *, p < 0.05, **, p < 0.01, N.S., not significant). (F-H) HSC- LX2 cells were stably transferred with ZFP36 Δ186-192 plasmid, ZFP36 plasmid, or FBXW7 plasmid, and then were treated with erastin (10 μM) for 24 h. The binding of ZFP36 and FBXW7, the ubiquitylation of ZFP36, and the protein levels of ZFP36 were determined (n = 3 in every group, *, p < 0.05, N.S., not significant). (I) HSC-LX2 cells were treated with erastin (10 μM) with or without autophagy inhibitors (3-MA, 10 mM; bafilomycin A1, 5 nM) for 24 h. The protein levels of ZFP36 were determined (n = 3 in every group, ***, p < 0.001, N.S., not significant).

Article Snippet: ZFP36 shRNA plasmid (sc-36760-SH, sc-36761-SH), FBXW7 shRNA plasmid (sc-37547-SH, sc-37548-SH) and empty vector were purchased from Santa Cruz Biotechnology.

Techniques: Ubiquitin Proteomics, Expressing, Sequencing, Binding Assay, Immunoprecipitation, Stable Transfection, Plasmid Preparation

Journal: Autophagy

Article Title: RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells.

doi: 10.1080/15548627.2019.1687985

Figure Lengend Snippet: Figure 4. Reduced ferroptosis by ZFP36 plasmid is associated with autophagy inactivation. (A) HSC-LX2 cells overexpressing ZFP36 were treated with sorafenib (10 μM) for 24 h, and total RNAs were extracted for RNA-Seq. Microarray heat map demonstrates clustering of HSC-LX2 cells. Hierarchical cluster analyses of significantly differentially expressed mRNAs: bright blue, underexpression; gray, no change; bright red, overexpression (Control vector, n = 3; ZFP36 plasmid, n = 3). (B-D) HSC-LX2 cells overexpressing ZFP36 and FBXW7 were treated with erastin (10 μM), sorafenib (10 μM), and RSL3 (2.5 μM) for 24 h. The protein levels of ATG3, ATG4A, ATG12– ATG5, BECN1, ATG7, ATG9A, and ATG16L1 were determined (n = 3 in every group). (E) HSC-LX2 cells overexpressing ZFP36 and FBXW7 were treated with sorafenib (10 μM) for 24 h. The protein complex of ATG12–ATG5-ATG16L1 was assayed by immunocoprecipitation (n = 3 in every group). (F) HSC-LX2 and HSC-T6 cells overexpressing ZFP36 and FBXW7 were treated with sorafenib (10 μM) for 24 h. The protein levels of LC3-I/II were determined (n = 3 in every group).

Article Snippet: ZFP36 shRNA plasmid (sc-36760-SH, sc-36761-SH), FBXW7 shRNA plasmid (sc-37547-SH, sc-37548-SH) and empty vector were purchased from Santa Cruz Biotechnology.

Techniques: Plasmid Preparation, RNA Sequencing, Microarray, Over Expression, Control

Journal: Autophagy

Article Title: RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells.

doi: 10.1080/15548627.2019.1687985

Figure Lengend Snippet: Figure 5. ZFP36 plasmid inhibits HSC ferroptosis by regulating autophagy signaling. (A) HSC-LX2 cells overexpressing ZFP36 and FBXW7 were stably transferred with pGM-CMV-GFP-hLC3 plasmid, and then were treated with sorafenib (10 μM) for 24 h. The green fluorescence spots were detected. Scale bars: 50 μm. Representative photographs were shown (n = 3 in every group, **, p < 0.01, ***, p < 0.001). (B) HSC-LX2 cells overexpressing ZFP36 and FBXW7 were treated with sorafenib (10 μM) for 24 h. The endogenous LC3 levels were determined by immunofluorescence. Scale bars: 50 μm. Representative photographs were shown (n = 3 in every group, **, p < 0.01, ***, p < 0.001). (C) HSC-LX2 cells were stably transferred with CMV-TurboRFP-EGFP-LC3-PGK-Puro plasmid and ZFP36 plasmid, and then were treated with sorafenib (10 μM) with or without autophagy inhibitors (3-MA, 10 mM; bafilomycin A1, 5 nM) for 24 h. The yellow fluorescence and red fluorescence spots were detected. Scale bars: 50 μm. Representative photographs were shown (n = 3 in every group, *, p < 0.05, **, p < 0.01). (D) HSC-LX2 cells overexpressing ZFP36 and FBXW7 were treated with sorafenib (10 μM) for 24 h. Autophagosomes and autolysosomes were determined by transmission electron microscopy. Scale bars: 0.5 μm. Representative photographs were shown (n = 3 in every group, *, p < 0.05, **, p < 0.01, ##, p < 0.01).

Article Snippet: ZFP36 shRNA plasmid (sc-36760-SH, sc-36761-SH), FBXW7 shRNA plasmid (sc-37547-SH, sc-37548-SH) and empty vector were purchased from Santa Cruz Biotechnology.

Techniques: Plasmid Preparation, Stable Transfection, Fluorescence, Immunofluorescence, Transmission Assay, Electron Microscopy

Journal: Autophagy

Article Title: RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells.

doi: 10.1080/15548627.2019.1687985

Figure Lengend Snippet: Figure 7. ZFP36 plasmid promotes autophagy inactivation and ATG16L1 mRNA decay via binding to the AU-rich elements. (A) The predicted hits of the ZFP36 signature motif in human ATG16L1 and TNF mRNA 3ʹ-UTR were assayed. (B) HSC-LX2 cells overexpressing ZFP36 were treated with sorafenib (10 μM), ActD (10 μg/ml) and DBR (5 μM) for indicated times. The remaining ATG16L1 mRNA levels were measured by real-time PCR and normalized to the results at 0 min after ActD/DRB treatment (n = 3 in every group, **, p < 0.01). (C) HSC-LX2 cells overexpressing ZFP36 were treated with sorafenib (10 μM) and CHX (20 μg/ml) for 150 min. ATG16L1 protein level was determined at the indicated time points (n = 3 in every group). (D) Association of endogenous ZFP36 with endogenous ATG16L1 mRNA was measured by real-time PCR after ribonucleoprotein immunoprecipitation (RNP IP) (n = 3 in every group, **, p < 0.01). (E) mRNA affinity isolation assay was performed with biotinylated transcripts of the ATG16L1 mRNA 5ʹ-UTR, coding region (CR), 3ʹ-UTR or the TNF mRNA 3ʹ-UTR (n = 3 in every group). (F) Luciferase constructs carrying the 3ʹ-UTRs of genes encoding ACTB (3ʹ-UTR-ACTB), TNF (3ʹ-UTR-TNF), ATG16L1 (3ʹ-UTR-ATG16L1) or empty vector (pGL3) was stably transfected into HSC-LX2 cells. Cells were treated with sorafenib (10 μM) for 24 h, and luciferase activities were measured and normalized to the activities obtained in pGL3-transfected cells without treatment (n = 3 in every group, ***, p < 0.001). (G) Luciferase constructs carrying the 3ʹ-UTRs of genes encoding ACTB (3ʹ-UTR-ACTB) and ATG16L1 (3ʹ-UTR- ATG16L1) were stably transfected into HSC-LX2 cells. Luciferase mRNA half-life was measured by real-time PCR after sorafenib (10 μM) and ActD/DRB treatment for indicated times (n = 3 in every group, ***, p < 0.001). (H) HSC-LX2 cells overexpressing ZFP36 were stably transfected with the luciferase constructs carrying the 3ʹ- UTRs of genes encoding ACTB (3ʹ-UTR-ACTB), TNF (3ʹ-UTR-TNF), ATG16L1 (3ʹ-UTR-ATG16L1) or empty vector (pGL3), and then were treated with sorafenib (10 μM) for 24 h. Luciferase activities were measured and normalized to the activities obtained in pGL3-transfected cells without treatment (n = 3 in every group, ***, p < 0.001). (I) ATG16L1 3ʹ-UTR luciferase construct was co-transfected with different amounts of ZFP36 plasmid into HSC-LX2 cells. Luciferase activity was detected after sorafenib (10 μM) treatment for 24 h and normalized to the cells transfected with empty plasmid (n = 3 in every group, *, p < 0.05, **, p < 0.01, ***, p < 0.001). (J) mRNA pull down assay was performed by mixing ATG16L1-AGE-Bio, TNF-AGE-Bio, and Mut-ATG16L1-AGE-Bio with total cell extracts from HSC-LX2 cells. Precipitates were prepared for western blot (n = 3 in every group). (K) Cold ATG16L1-AGE probes or Mut-ATG16L1-AGE probes with different concentrations were used to compete for the binding between ATG16L1-AGE-Bio and ZFP36 (n = 3 in every group).

Article Snippet: ZFP36 shRNA plasmid (sc-36760-SH, sc-36761-SH), FBXW7 shRNA plasmid (sc-37547-SH, sc-37548-SH) and empty vector were purchased from Santa Cruz Biotechnology.

Techniques: Plasmid Preparation, Binding Assay, Real-time Polymerase Chain Reaction, Immunoprecipitation, Isolation, Luciferase, Construct, Stable Transfection, Transfection, Activity Assay, Pull Down Assay, Western Blot

Journal: Autophagy

Article Title: RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells.

doi: 10.1080/15548627.2019.1687985

Figure Lengend Snippet: Figure 10. RNA-binding protein ZFP36 protects against ferroptosis by regulating autophagy signaling pathway in HSCs. ZFP36 overexpression can result in ATG16L1 mRNA decay via binding to the AREs in the 3ʹ-UTR, thus triggering autophagy inactivation, blocking autophagic ferritin degradation, and eventually conferring resistance to ferroptosis.

Article Snippet: ZFP36 shRNA plasmid (sc-36760-SH, sc-36761-SH), FBXW7 shRNA plasmid (sc-37547-SH, sc-37548-SH) and empty vector were purchased from Santa Cruz Biotechnology.

Techniques: RNA Binding Assay, Over Expression, Binding Assay, Blocking Assay

Journal: Frontiers in Microbiology

Article Title: Impact of glucokinase activators on the gut microbiota of high-fat diet-induced obese and type 2 diabetic mice

doi: 10.3389/fmicb.2026.1783385

Figure Lengend Snippet: (A) Experimental design; (B) body weight; (C) FBG levels before treatment started (at 5 weeks); (D) FBG levels after treatment (at 9 weeks). All data are presented as mean ± SE (CD_Vehicle1 group: n = 7, CD_Vehicle2 group: n = 6, HFD_Vehicle1 group: n = 9, HFD_Vehicle2 group: n = 9, HFD_Dorzagliatin group: n = 9, HFD_TTP399 group: n = 9). One-way or two-way ANOVA followed by Tukey’s multiple comparison test was used to determine significant differences. **** p < 0.0001 (CD_Vehicle1 vs. HFD_Vehicle1, CD_Vehicle1 vs. HFD_Dorzagliatin), ** p < 0.01 (HFD_Vehicle1 vs. HFD_Dorzagliatin), #### p < 0.0001 (CD_Vehicle2 vs. HFD_Vehicle2), ### p < 0.001 (CD_Vehicle2 vs. HFD_TTP399).

Article Snippet: Dorzagliatin (HY-109030) and TTP399 (HY-147254) were purchased from MedChemExpress (MCE, United States).

Techniques: Comparison

Journal: Frontiers in Microbiology

Article Title: Impact of glucokinase activators on the gut microbiota of high-fat diet-induced obese and type 2 diabetic mice

doi: 10.3389/fmicb.2026.1783385

Figure Lengend Snippet: Effects of dorzagliatin and TTP399 on glucose tolerance and insulin tolerance in HFD-fed mice. (A) OGTT after 3 weeks of treatment; (B) AUC calculated from the glucose concentrations versus time for the period of 0–120 min after glucose load by the trapezoidal rule; (C) Blood glucose levels at 30 min post-administration during the OGTT; (D) ITT after 4 weeks of treatment; (E) AUC of the ITT; (F) Fasting insulin (FINS) levels and HOMA-IR value (G,H) during the ITT. Values are expressed as mean ± SE (CD_Vehicle1 group: n = 7, CD_Vehicle2 group: n = 6, HFD_Vehicle1 group: n = 9, HFD_Vehicle2 group: n = 9, HFD_Dorzagliatin group: n = 9, HFD_TTP399 group: n = 9). Differences were assessed by one-way ANOVA or two-way ANOVA. **** p < 0.0001 (HFD_Vehicle1 vs. HFD_Dorzagliatin), *** p < 0.001 (E: HFD_Vehicle1 vs. HFD_Dorzagliatin, H: CD_Vehicle1 vs. HFD_Vehicle1), ** p < 0.01 (HFD_Vehicle1 vs. HFD_Dorzagliatin), * p < 0.05 (HFD_Vehicle1 vs. HFD_Dorzagliatin), ## p < 0.01(HFD_Vehicle2 vs. HFD_TTP399).

Article Snippet: Dorzagliatin (HY-109030) and TTP399 (HY-147254) were purchased from MedChemExpress (MCE, United States).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Impact of glucokinase activators on the gut microbiota of high-fat diet-induced obese and type 2 diabetic mice

doi: 10.3389/fmicb.2026.1783385

Figure Lengend Snippet: Effects of dorzagliatin on the composition of the gut microbiota in HFD-fed mice. (A) Alpha diversity analysis of observed-features index and Chao1 index were analyzed using the Kruskal-Wallis test; (B) Relative abundance of the gut microbiota at the phylum levels; (C) NMDS analysis plot of the gut microbiota based on unweighted-Unifrac distance; (D) Relative abundance of the gut microbiota at the genus levels. Values are expressed as mean ± SE (CD_Vehicle1 group: n = 7, CD_Vehicle2 group: n = 6, HFD_Vehicle1 group: n = 9, HFD_Vehicle2 group: n = 9, HFD_Dorzagliatin group: n = 9, HFD_TTP399 group: n = 9). Differences were assessed by one-way ANOVA.

Article Snippet: Dorzagliatin (HY-109030) and TTP399 (HY-147254) were purchased from MedChemExpress (MCE, United States).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Impact of glucokinase activators on the gut microbiota of high-fat diet-induced obese and type 2 diabetic mice

doi: 10.3389/fmicb.2026.1783385

Figure Lengend Snippet: Effects of TTP399 on the composition of the gut microbiota in HFD-fed mice. (A) Alpha diversity analysis of observed-features index and Chao1 index were analyzed using the Kruskal-Wallis test; (B) Relative abundance of the gut microbiota at the phylum levels; (C) PCoA analysis plot of the gut microbiota based on Jaccard distances; (D) Relative abundance of the gut microbiota at the genus levels. Values are expressed as mean ± SE (CD_Vehicle1 group: n = 7, CD_Vehicle2 group: n = 6, HFD_Vehicle1 group: n = 9, HFD_Vehicle2 group: n = 9, HFD_Dorzagliatin group: n = 9, HFD_TTP399 group: n = 9). Differences were assessed by one-way ANOVA. * p < 0.05.

Article Snippet: Dorzagliatin (HY-109030) and TTP399 (HY-147254) were purchased from MedChemExpress (MCE, United States).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Impact of glucokinase activators on the gut microbiota of high-fat diet-induced obese and type 2 diabetic mice

doi: 10.3389/fmicb.2026.1783385

Figure Lengend Snippet: Dorzagliatin and TTP399 have no significant effect on intestinal barrier integrity or inflammation in HFD-fed mice. (A) HE staining results of ileum; (B) HE staining results of colon; (C) Expression levels of Tjp1, claudin-1, and occludin in the duodenum, jejunum, ileum, and colon after 4 weeks of treatment; (D) Expression levels of TNF- α , IL-1β, and IL-6 in the duodenum, jejunum, ileum, and colon after 4 weeks of treatment. Values are expressed as mean ± SE (CD_Vehicle1 group: n = 7, CD_Vehicle2 group: n = 6, HFD_Vehicle1 group: n = 9, HFD_Vehicle2 group: n = 9, HFD_Dorzagliatin group: n = 9, HFD_TTP399 group: n = 9). Differences were assessed by one-way ANOVA.

Article Snippet: Dorzagliatin (HY-109030) and TTP399 (HY-147254) were purchased from MedChemExpress (MCE, United States).

Techniques: Staining, Expressing

Journal: Frontiers in Microbiology

Article Title: Impact of glucokinase activators on the gut microbiota of high-fat diet-induced obese and type 2 diabetic mice

doi: 10.3389/fmicb.2026.1783385

Figure Lengend Snippet: Correlation analysis. (A) Correlation analysis between biochemical indices, inflammatory markers, and gut microbiota; (B) Correlation analysis between intestinal barrier integrity and gut microbiota. (CD_Vehicle1 group: n = 7, CD_Vehicle2 group: n = 6, HFD_Vehicle1 group: n = 9, HFD_Vehicle2 group: n = 9, HFD_Dorzagliatin group: n = 9, HFD_TTP399 group: n = 9).

Article Snippet: Dorzagliatin (HY-109030) and TTP399 (HY-147254) were purchased from MedChemExpress (MCE, United States).

Techniques: