Journal: Molecular Medicine Reports

Article Title: lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating β-catenin via enhancing H3K18 lactylation

doi: 10.3892/mmr.2025.13763

Figure Lengend Snippet: Long non-coding RNA NEAT1 regulates β-catenin by positively affecting the lactyltransferases AARS1 and P300 in hemangioma cells. (A) The knockdown efficiencies of AARS1, AARS2, CBP, KAT5, KAT8 and P300 were confirmed by RT-qPCR; ****P<0.0001 vs. NC (n=3). (B) After AARS1, AARS2, CBP, KAT5, KAT8 and P300 were knocked down, the CTNNB1 mRNA level was detected using RT-qPCR; *P<0.05, ***P<0.001, ****P<0.0001 vs. NC (n=3). (C) The knockdown efficiencies of SIRT1, SIRT2, SIRT3, HDAC1, HDAC2 and HDAC3 were confirmed by RT-qPCR; ****P<0.0001 vs. NC (n=3). (D) After SIRT1, SIRT2, SIRT3, HDAC1, HDAC2 and HDAC3 were knocked down, the CTNNB1 mRNA level was detected using RT-qPCR; ****P<0.0001 vs. NC (n=3). (E) The knockdown efficiency of AARS1 and P300 was confirmed by western blotting (n=3). (F) Semi-quantitative analysis of the levels of AARS1 and P300 in AARS1-knock down and P300-knock down cells detected by western blotting; **P<0.01, ****P<0.0001 vs. NC (n=3). (G) The protein level of β-catenin after AARS1 and P300 knockdown was detected using western blotting (n=3). (H) Semi-quantitative analysis of the levels of β-catenin in AARS1-knock down and P300-knock down cells detected by western blotting; **P<0.01, ***P<0.001 vs. NC (n=3). (I) The mRNA levels of AARS1 and P300 after NEAT1 knockdown were detected using RT-qPCR; ****P<0.0001 vs. NC (n=3). (J) The protein levels of AARS1 and P300 after NEAT1 knockdown were detected using western blotting (n=3). (K) Semi-quantitative analysis of the levels of AARS1 and P300 in NEAT1-knock down cells detected by western blotting; **P<0.01, ***P<0.001, ****P<0.0001 vs. NC (n=3). ns, not significant; NC, negative control; AARS, alany-tRNA synthetase; CBP, CREB binding lysine acetyltransferase; KAT, lysine acetyltransferase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; CTNNB1, β-catenin gene; SIRT, sirtuin; HDAC, histone deacetylase; siRNA, small interfering RNA.

Article Snippet: The primary antibodies used were LDHB (cat. no. 14824-1-AP; 1:5,000), β-catenin (cat. no. 51067-2-AP; 1:5,000), alanyl-tRNA synthetase 1 (AARS1; cat. no. 17394-1-AP; 1:2,000), P300 (cat. no. 20695-1-AP; 1:1,000) and GAPDH (cat. no. 60004-1-Ig; 1:50,000) from Proteintech Group, Inc., and L-lactyl-Histone H3 (Lys18) (cat. no. PTM-1406RM; 1:1,000), L-lactyl lysine (pan-lac; cat. no. PTM-1401RM; 1:1,000) and Histone H3 (cat. no. PTM-1002RM; 1:1,000) from PTM BIO LLC.

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Negative Control, Binding Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Histone Deacetylase Assay, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating β-catenin via enhancing H3K18 lactylation

doi: 10.3892/mmr.2025.13763

Figure Lengend Snippet: β-catenin and H3K18 lactylation levels were elevated in IH tissues. (A) The viability of hemangioma cells treated with a P300 inhibitor (A-485) were measured via Cell Counting Kit-8 assay; ***P<0.001 (n=3). (B) The colony formation ability of hemangioma cells treated with a P300 inhibitor (A-485) were measured via colony formation assays (n=3). (C) Quantitative analysis of the colony formation results; **P<0.01 (n=3). (D) The CTNNB1 mRNA level after A-485 treatment was detected using RT-qPCR; **P<0.01 (n=3). (E) The protein level of β-catenin after A-485 treatment was detected using western blotting (n=3). (F) Semi-quantitative analysis of the levels of β-catenin in A-485-treated cells detected by western blotting; **P<0.01 (n=3). (G) The CTNNB1 mRNA level in IH tissues was detected using RT-qPCR; **P<0.01 (n=10). (H) The level of β-catenin protein in IH tissues was detected using western blotting. (I) The level of H3K18 lactylation in IH tissues was detected using western blotting. (J) Proposed model of long non-coding RNA NEAT1 regulation of the LDHB/H3K18-lactylation/β-catenin signaling cascade. IH, infantile hemangioma; NC, negative control; CTNNB1, β-catenin gene; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; AARS1, alany-tRNA synthetase 1; LDHB, lactate dehydrogenase B.

Article Snippet: The primary antibodies used were LDHB (cat. no. 14824-1-AP; 1:5,000), β-catenin (cat. no. 51067-2-AP; 1:5,000), alanyl-tRNA synthetase 1 (AARS1; cat. no. 17394-1-AP; 1:2,000), P300 (cat. no. 20695-1-AP; 1:1,000) and GAPDH (cat. no. 60004-1-Ig; 1:50,000) from Proteintech Group, Inc., and L-lactyl-Histone H3 (Lys18) (cat. no. PTM-1406RM; 1:1,000), L-lactyl lysine (pan-lac; cat. no. PTM-1401RM; 1:1,000) and Histone H3 (cat. no. PTM-1002RM; 1:1,000) from PTM BIO LLC.

Techniques: Cell Counting, Quantitative RT-PCR, Western Blot, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction