Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p was decreased in the livers of MAFLD mice and FFA-induced HepG2 cells. (A) MiR-300-3p was decreased in the livers of MAFLD mice. (B) MiR-300-3p was downregulated in the FFA-induced HepG2 cells. (C) MiR-300-3p was downregulated in HepG2 cells successfully. (D) MiR-300-3p was overexpressed in HepG2 cells successfully. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01. NC, Healthy control; OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Control, Over Expression, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: Downregulation of miR-300-3p promotes FFA-induced lipid accumulation and hepatic inflammation in HepG2 cells. (A) The experimental flow of the MAFLD cell model construction and miR-300-3p mimics, miR-300-3p inhibitors and as well as corresponding scrambled controls transfection in HepG2 cells. (B,C) Intercellular TG contents in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (D) Oil Red O staining (400×) and the relative areas of lipid droplets in miR-300-3p-inhibited and -overexpressing in HepG2 cells. (E,F) mRNA levels of FASN, SREBP-1c, IL-6, IL-2 and TNF-αin miR-300-3p -inhibited and -overexpressing in HepG2 cells. (G) Protein levels of FASN, SREBP-1c and TNF-α in miR-300-3p -inhibited and -overexpressing in HepG2 cells. Data in (B,C) and (E–G) are means±SDs (n = 3). *P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001. OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Transfection, Staining, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: Downregulation of miR-300-3p induces apoptosis in HepG2 cells to promote it. (A,B) The apoptosis rate in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (C) mRNA levels of Bax, Bcl-2 and Caspase-3 in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (D) Protein levels of Bax, Bcl-2 and Caspase-3 in miR-300-3p -overexpressing and -inhibited in HepG2 cells. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p regulates STX17 in HepG2 cells. (A,B) GO enrichment analysis (Biological Process category) and KEGG enrichment analysis diagrams of sixteen predicted target genes of miR-300-3p. (C) The predicted binding sites between miR-300-3p and STX17 were putatively identified via the TargetScan database. (D) Dual-luciferase reporter assay (DLRA) in HepG2 cells validated that STX17 is a direct target gene of miR-300-3p. (E,F) The mRNA and protein expression levels of STX17 in HepG2 cells with miR-300-3p inhibition or overexpression. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Inhibition, Over Expression, Control

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: P62 and LC3-II in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (A) The protein levels of P62 and LC3-II in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (B) P62 and LC3-II in miR-300-3p -inhibited and -overexpressing by Immunofluorescence assay. Data in (A-D) are means±SDs (n = 3). *P < 0.05, **P < 0.01,***P < 0.001,****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p promoted autophagy by regulating STX17, reduces lipid accumulation, inflammatory response, and decreases hepatocyte apoptosis. (A) STX17 was inhibited or overexpressed in HepG2 cells successfully. (B) Protein levels of FASN, SREBP-1c and TNF-αin STX17 -inhibited and -overexpressing in HepG2 cells. (C,D) Intercellular TG contents in STX17-inhibited and -overexpressing in HepG2 cells. (E) Oil Red O staining (400×) and relative areas of lipid droplets in STX17-inhibited and -STX17 in HepG2 cells. (F) Protein levels of Bax, Capase-3 and Bcl-2 in STX17 -inhibited and -overexpressing in HepG2 cells. Data in (A–D) and (F) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, miR-300-3p overexpression control; IC,miR-300-3p inhibition control; Control, STX17-overexpression control or STX17-inhibition control; Si-STX17, STX17-inhibition; Over-STX17, STX17-overexpression.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Staining, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: P62, LC3-II in STX17-inhibited and -overexpressing in HepG2 cells. (A) Protein levels of P62, LC3-II in STX17-inhibited and -overexpressing in HepG2 cells. (B) P62 and LC3-II in STX17-inhibited and -overexpressing by Immunofluorescence assay. Data in (A-D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Over Expression, Control, Inhibition

Journal: Biotechnology Reports

Article Title: Insulin Resistance, Anti-inflammatory, and Antioxidant In vitro Activity of Heterologously Expressed Arenin from Dryophytes arenicolor

doi: 10.1016/j.btre.2026.e00947

Figure Lengend Snippet: In vitro modulation of hepatic insulin resistance by arenin in hepatic insulin-resistant (HepG2) cells . Arenin concentrations (3.90–1000 µg/mL) were incubated for 5 h in hepatic insulin-resistant cells. Negative control (N) cells were cultured in the absence of insulin, whereas insulin-resistant (IR) control cells were cultured in the presence of insulin. Rosiglitazone (ROSI) at 10 μM (3.57 µg/mL) was used as the positive reference control. Different letters abc indicate statistical differences between concentrations analyzed using Tukey test ( p < 0.05).

Article Snippet: Human hepatocyte carcinoma (HepG2) cells, human primary dermal fibroblasts (HDFa) cells, and murine macrophage (RAW 264.7) cells were obtained from the American Type Culture Collection (ATCC, VA, USA).

Techniques: In Vitro, Incubation, Negative Control, Cell Culture, Control

Journal: bioRxiv

Article Title: On the role of cell chaining in the attenuation of a Listeria monocytogenes divIVA mutant

doi: 10.64898/2026.01.05.697648

Figure Lengend Snippet: (A) Contribution of divIVA , mreB and ezrA to hepatocyte invasion. Invasion of L. monocytogenes strains EGD-e (wt), BUG2214 (Δ prfA ), LMS2 (Δ divIVA ), LMSW49 (i mreB ) and LMJR183 (i ezrA ) pre-grown in the absence or presence of 1 mM IPTG into HepG2 hepatocytes. Values are expressed relative to wild type. Average values and standard deviations are shown. Asterisks mark statistically significant differences compared to wild type ( P <0.01, t -test with Bonferroni Holm correction). (B) Contribution of divIVA , mreB and ezrA to intracellular growth in macrophages. J774 mouse macrophages were infected with L. monocytogenes strains EGD-e (wt), LMS250 (Δ hly ), LMS2 (Δ divIVA ), LMSW49 (i mreB ) and LMJR183 (i ezrA ) and then cultivated in the absence or presence of IPTG, where appropriate. Average values and standard deviations were calculated from experiments performed in triplicates. Statistically significant differences (compared to wild type or between depleted and induced conditions) are marked by asterisks ( P <0.01, t -test with Bonferroni-Holm correction, n. s. – not significant). (C) Effect of DivIVA, MreB or EzrA depletion on cell-to-cell spread. Plaque formation assay using 3T3 mouse fibroblasts with L. monocyctogenes strains EGD-e (wt), LMS30 (i divIVA ), LMSW49 (i mreB ) and LMJR183 (i ezrA ) ± IPTG. (D) Necessity of DivIVA, MreB and EzrA for actin coating and actin tail formation. J774 mouse macrophages were infected with DsRed producing L. monocytogenes strains LMJD20 (wt), LMSF2 (Δ actA ), LMSW205 (Δ divIVA ), LMSW202 (EzrA depletion) and LMSW203 (MreB depletion). Infections were carried out in the absence of IPTG and analyzed by fluorescence microscopy 4 hours post infection after DAPI and phalloidin staining. Composite images are shown.

Article Snippet: Invasion of L. monocytogenes into HepG2 human hepatocytes (ATCC® HB-8065 TM ) was quantified as described earlier [ ].

Techniques: Infection, Plaque Formation Assay, Fluorescence, Microscopy, Staining

Journal: bioRxiv

Article Title: On the role of cell chaining in the attenuation of a Listeria monocytogenes divIVA mutant

doi: 10.64898/2026.01.05.697648

Figure Lengend Snippet: (A) The prfA* mutation suppresses the invasion defect of the Δ divIVA mutant. Invasion of L. monocytogenes strains EGD-e (wt), BUG2214 (Δ prfA ), BUG3057 ( prfA* ), LMS2 (Δ divIVA ) and LMSW251 (Δ divIVA prfA* ) into HepG2 hepatocytes. Values are expressed relative to wild type. Average values and standard deviations are shown. The asterisks mark statistically significant differences compared to wild type ( P <0.01, t -test with Bonferroni-Holm correction) or to the Δ divIVA prfA* mutant ( P <0.01, t -test, n. s. – not significant). (B) No suppression of the intracellular replication defect of the Δ divIVA mutant by the prfA* mutation. J774 mouse macrophages were infected with the same strains as in panel A. Average values and standard deviations were calculated from experiments performed in triplicates. No statistically significant difference in the number of intracellular bacteria was detected between Δ divIVA and Δ divIVA prfA* cells six hours post infection ( P <0.01, t -test, n. s. – not significant). (C) Dominance of the Δ divIVA deletion over the prfA* mutation in a plaque forming assay. The same strains as above were used to infect 3T3 mouse fibroblasts to analyze cell-to-cell spread.

Article Snippet: Invasion of L. monocytogenes into HepG2 human hepatocytes (ATCC® HB-8065 TM ) was quantified as described earlier [ ].

Techniques: Mutagenesis, Infection, Bacteria

Journal: bioRxiv

Article Title: On the role of cell chaining in the attenuation of a Listeria monocytogenes divIVA mutant

doi: 10.64898/2026.01.05.697648

Figure Lengend Snippet: (A) Localisation of Δ divIVA suppressor mutations M95I and T123A in the SecA2 protein. SecA2 domains (colored) and walker A and B motifs (black) are indicated. Abbreviations: NBD –nucleotide binding domain, PPXD – preprotein crosslinking domain, IRA 1/2 – intramolecular regulator of ATPase domains 1 and 2, SD – scaffold domain. (B) Micrographs showing the cellular morphology of L. monocytogenes strains EGD-e (wt), LMS2 (Δ divIVA ), LMSW222 (Δ divIVA secA2 M95I ) and LMSW223 (Δ divIVA secA2 T123A ) during growth in BHI broth at 37°C. Membranes were stained with nile red. Scale bar is 2 µm. (C) Separation of secretome samples of the same set of strains by SDS-PAGE. The position of p60 (CwhA) is indicated. (D) Flagellar motility of the same set of strains on swarming agar after two days of incubation at 30°C. (E) Cell-to-cell spread of the same strains in 3T3 mouse fibroblast. (F) Full suppression of the Δ divIVA invasion defect by secA2 mutations. The same set of strains as in panel A was used to infect HepG2 hepatocytes. Values are expressed relative to wild type. Average values and standard deviations are shown. The asterisk marks a statistically significant difference ( P <0.01, t -test with Bonferroni-Holm correction, n. s. – not significant). (G) Partial suppression of the intracellular replication defect of the Δ divIVA mutant by secA2 mutations. J774 mouse macrophages were infected with the same strains as above and intracellular bacteria were enumerated right after infection (0 h p. i.) and six hours later (6 h p. i.). Average values and standard deviations were calculated from experiments performed in triplicates. Statistically significant differences are indicated by asterisks ( P <0.01, t -test with Bonferroni-Holm correction).

Article Snippet: Invasion of L. monocytogenes into HepG2 human hepatocytes (ATCC® HB-8065 TM ) was quantified as described earlier [ ].

Techniques: Binding Assay, Staining, SDS Page, Incubation, Mutagenesis, Infection, Bacteria