medium cm raw 264 7 cells  (ATCC)

Journal: Frontiers in Oncology

Article Title: Emodin inhibits colon cancer tumor growth by suppressing tumor cell glycolysis through inhibition of NAT10-mediated PGK1 ac4C modification

doi: 10.3389/fonc.2025.1575391

Figure Lengend Snippet: Emodin inhibited cell viability of colon cancer cells. (A) The molecular structure of emodin. (B, C) Cell viability of HCT-116 and SW480 cells was measured by MTT assay. N = 3 per group. Data analysis was performed using ANOVA.

Article Snippet: Extracellular acidification rate (ECAR) of HCT-116 and SW480 cells was measured using an ECAR fluorometric assay kit (Elabscience, Wuhan, China).

Techniques: MTT Assay

Journal: Frontiers in Oncology

Article Title: Emodin inhibits colon cancer tumor growth by suppressing tumor cell glycolysis through inhibition of NAT10-mediated PGK1 ac4C modification

doi: 10.3389/fonc.2025.1575391

Figure Lengend Snippet: Emodin inhibited cell proliferation and glycolysis in colon cancer cells. (A, B) Cell proliferation was evaluated by EdU staining. (C) Glucose uptake of HCT-116 and SW480 cells was measured using a glucose uptake assay kit. (D) Lactate production was evaluated using a lactate content detection kit. (E, F) Real-time cell metabolism was assessed by measuring ECAR. N = 3 per group. Data analysis was performed using ANOVA.

Article Snippet: Extracellular acidification rate (ECAR) of HCT-116 and SW480 cells was measured using an ECAR fluorometric assay kit (Elabscience, Wuhan, China).

Techniques: Staining

Journal: Frontiers in Oncology

Article Title: Emodin inhibits colon cancer tumor growth by suppressing tumor cell glycolysis through inhibition of NAT10-mediated PGK1 ac4C modification

doi: 10.3389/fonc.2025.1575391

Figure Lengend Snippet: Emodin suppressed ac4C modification by inhibiting NAT10 expression in colon cancer cells. (A, B) The ac4C levels of HCT-116 and SW480 cells were detected by dot blot assay. (C) The expression of NAT10 in HCT-116 and SW480 cells was measured by qPCR. (D) The protein levels of NAT10 in HCT-116 and SW480 cells were detected by western blot. The interaction between NAT10 and eomdin was determined by (E) molecular docking and (F) SRP assay. N = 3 per group. Data analysis was performed using student’s t-test.

Article Snippet: Extracellular acidification rate (ECAR) of HCT-116 and SW480 cells was measured using an ECAR fluorometric assay kit (Elabscience, Wuhan, China).

Techniques: Modification, Expressing, Dot Blot, Western Blot

Journal: Frontiers in Oncology

Article Title: Emodin inhibits colon cancer tumor growth by suppressing tumor cell glycolysis through inhibition of NAT10-mediated PGK1 ac4C modification

doi: 10.3389/fonc.2025.1575391

Figure Lengend Snippet: NAT10 overexpression restored glycolysis inhibited by emodin in colon cancer cells. (A) The expression of NAT10 was measured by qPCR. (B) The protein levels of NAT10 were detected by western blot. (C, D) Cell proliferation of HCT-116 and SW480 cells was evaluated by EdU staining. (E) Glucose uptake was evaluated using a glucose uptake assay kit. (F) Lactate production was evaluated using a lactate content detection kit. (G) Real-time cell metabolism was assessed by measuring ECAR. (H) The protein levels of HK2, PFK1 and LDHA were detected by western blot. N = 3 per group. Data analysis was performed using student’s t-test (A) or ANOVA (D–G) .

Article Snippet: Extracellular acidification rate (ECAR) of HCT-116 and SW480 cells was measured using an ECAR fluorometric assay kit (Elabscience, Wuhan, China).

Techniques: Over Expression, Expressing, Western Blot, Staining

Journal: Frontiers in Oncology

Article Title: Emodin inhibits colon cancer tumor growth by suppressing tumor cell glycolysis through inhibition of NAT10-mediated PGK1 ac4C modification

doi: 10.3389/fonc.2025.1575391

Figure Lengend Snippet: NAT10 knockdown suppressed PGK1 mRNA stability by inhibiting its ac4C modification. (A) The expression of NAT10 was measured by qPCR. (B) The protein levels of NAT10 were detected by western blot. (C) The expression of PGK1 was measured by qPCR. (D) The protein levels of PGK1 were detected by western blot. (E) The ac4C levels of HCT-116 and SW480 cells were detected by MeRIP. (F) The ac4C modification site of PGK1 was predicted using the PACES database. (G) The interaction between NAT10 and PGK1 was identified by RIP. (H) The luciferase activity of WT-PGK1 and MUT-PGK1 of HCT-116 cells. (I) The stability of PGK1 mRNA was measured by qPCR after HCT-116 cells treated with 5 μg/mL actinomycin D for 0, 4, 8, and 12 h. N = 3 per group. Data analysis was performed using student’s t-test (A, C, D, G) or ANOVA (F, H) .

Article Snippet: Extracellular acidification rate (ECAR) of HCT-116 and SW480 cells was measured using an ECAR fluorometric assay kit (Elabscience, Wuhan, China).

Techniques: Knockdown, Modification, Expressing, Western Blot, Luciferase, Activity Assay

Journal: Frontiers in Oncology

Article Title: Emodin inhibits colon cancer tumor growth by suppressing tumor cell glycolysis through inhibition of NAT10-mediated PGK1 ac4C modification

doi: 10.3389/fonc.2025.1575391

Figure Lengend Snippet: Cell proliferation and glycolysis of colon cancer cells inhibited by NAT10 knockdown was restored by PGK1 overexpression. (A) The expression of PGK1 was measured by qPCR. (B) The protein levels of PGK1 were detected by western blot. (C, D) Cell proliferation of HCT-116 and SW480 cells was evaluated by EdU staining. (E) Glucose uptake was evaluated using a glucose uptake assay kit. (F) Lactate production was evaluated using a lactate content detection kit. (G) Real-time cell metabolism was assessed by measuring ECAR. (H) The protein levels of HK2, PFK1 and LDHA were detected by western blot. N = 3 per group. Data analysis was performed using student’s t-test (A) or ANOVA (D–G) .

Article Snippet: Extracellular acidification rate (ECAR) of HCT-116 and SW480 cells was measured using an ECAR fluorometric assay kit (Elabscience, Wuhan, China).

Techniques: Knockdown, Over Expression, Expressing, Western Blot, Staining

Journal: Genes

Article Title: Transcriptional Activation of the TREM2 Gene by ZEB2 in a Zinc Finger-Dependent Manner

doi: 10.3390/genes16111329

Figure Lengend Snippet: The 5 kb upstream region and 5′-UTR of TREM2 recapitulate its cell type-specific expression. ( A ) Schematic representation of the reporter construct ( T2-5k-u-mCherry ). The CMV promoter in the mCherry-N3 vector was replaced with a fragment containing the 5 kb sequence upstream of the TREM2 transcription start site and its 5′-UTR. ( B ) T2-5k-u-mCherry was transfected into HEK293, THP-1, and HMC3 cells. Representative fluorescence images of mCherry expression are shown. Nuclei were counterstained with Hoechst33342. Scale bars, 50 μm. ( C ) Schematic diagram of the T2-5k-mCherry construct lacking the TREM2 5′-UTR. ( D ) Fluorescent images of THP-1 cells transfected with T2-5k-u-mCherry or T2-5k-mCherry . Scale bars, 50 μm. ( E ) Quantification of mCherry-positive cells relative to Hoechst-positive cells using the IN Cell Analyzer. Error bars represent SDs; n = 3; Welch’s t -test. ( F ) Fluorescence images of HMC3 cells transfected with T2-5k-u-mCherry or T2-5k-mCherry . Scale bars, 50 μm. ( G ) Quantification of mCherry-positive cells relative to Hoechst-positive cells analyzed as in ( E ). Error bars represent SDs; n = 3; Welch’s t -test.

Article Snippet: The HMC3 cell line (EP-CL-0620) was obtained from Elabscience (Houston, TX, USA).

Techniques: Expressing, Construct, Plasmid Preparation, Sequencing, Transfection, Fluorescence

Journal: Genes

Article Title: Transcriptional Activation of the TREM2 Gene by ZEB2 in a Zinc Finger-Dependent Manner

doi: 10.3390/genes16111329

Figure Lengend Snippet: ZEB2 overexpression or knockdown modulates TREM2 mRNA and protein expression. ( A ) RT-qPCR analysis of ZEB2 mRNA in doxycycline-treated inducible EGFP and EGFP-ZEB2 cells. Expression was normalized to ACTB . Error bars represent SDs; n = 3; Welch’s t -test. ( B ) TREM2 mRNA levels normalized to ACTB , quantified from the same samples as in ( A ). Error bars indicate SDs; n = 3; Welch’s t -test. ( C ) ZEB2 siRNAs were transfected into HMC3 cells. ZEB2 mRNA levels were measured by RT-qPCR and normalized to B2M . Error bars indicate SDs; n = 5; Tukey’s test. ( D ) TREM2 mRNA levels quantified as in ( C ). Error bars indicate SDs; n = 5; Tukey’s test. ( E ) Membrane-bound protein fractions from HMC3 cells were analyzed for TREM2 protein levels following ZEB2 knockdown. APP served as a loading control. ( F ) Quantification of ( E ). Error bars indicate SDs; n = 5; Welch’s t -test. ( G ) Relative YY1 mRNA expression in HMC3 cells transfected with YY1 siRNA. YY1 mRNA levels were normalized to B2M. Error bars indicate SDs; n = 3; Welch’s test. ( H ) TREM2 mRNA levels in HMC3 cells transfected with siZEB2 or siYY1. Error bars indicate SDs; n = 5; Tukey’s t-test. ( I ) Relative luciferase activity induced by EGFP-ZEB1. Error bars indicate SDs; n = 3; Welch’s test.

Article Snippet: The HMC3 cell line (EP-CL-0620) was obtained from Elabscience (Houston, TX, USA).

Techniques: Over Expression, Knockdown, Expressing, Quantitative RT-PCR, Transfection, Membrane, Control, Luciferase, Activity Assay