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Knocking down long non-coding RNA NEAT1 inhibits cellular protein lactylation and <t>downregulates</t> <t>β-catenin</t> in hemangioma cells. (A) Cellular lactate levels were measured after NEAT1 knockdown; ****P<0.0001 vs. NC (n=3). (B) The level of pan-lactylation in NEAT1-knock down cells were detected by western blotting (n=3). (C) H3K18 lactylation in NEAT1-knock down cells was detected by western blotting (n=3). (D) Quantitative analysis of the levels of H3K18la in the western blotting results; *P<0.05, **P<0.01 vs. NC (n=3). (E) The knockdown efficiency of <t>LDHB</t> was detected by western blotting; *P<0.05 (n=3). (F) The viability of LDHB-knock down cells was detected via Cell Counting Kit-8 assay; **P<0.01 (n=3). (G) Colony formation ability of LDHB-knock down cells was detected via a colony formation assay; **P<0.01 (n=3). (H) The mRNA levels of CTNNB1 in NEAT1-knock down cells were detected using RT-qPCR. ****P<0.0001 vs. NC (n=3). (I) The mRNA levels of CTNNB1 in LDHB-knock down cells were detected using RT-qPCR. ***P<0.001 vs. NC (n=3). (J) The protein levels of β-catenin in NEAT1-knock down and LDHB-knock down cells were measured using western blotting (n=3). (K) Semi-quantitative analysis of the levels of β-catenin in NEAT1-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). (L) Semi-quantitative analysis of the levels of β-catenin in LDHB-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). NC, negative control; H3K18la, Histone H3K18 lactylation; si, small interfering (RNA); LDHB, lactate dehydrogenase B; CTNNB1, β-catenin gene; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Ldhb, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating β-catenin via enhancing H3K18 lactylation"

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

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2025.13763

Knocking down long non-coding RNA NEAT1 inhibits cellular protein lactylation and downregulates β-catenin in hemangioma cells. (A) Cellular lactate levels were measured after NEAT1 knockdown; ****P<0.0001 vs. NC (n=3). (B) The level of pan-lactylation in NEAT1-knock down cells were detected by western blotting (n=3). (C) H3K18 lactylation in NEAT1-knock down cells was detected by western blotting (n=3). (D) Quantitative analysis of the levels of H3K18la in the western blotting results; *P<0.05, **P<0.01 vs. NC (n=3). (E) The knockdown efficiency of LDHB was detected by western blotting; *P<0.05 (n=3). (F) The viability of LDHB-knock down cells was detected via Cell Counting Kit-8 assay; **P<0.01 (n=3). (G) Colony formation ability of LDHB-knock down cells was detected via a colony formation assay; **P<0.01 (n=3). (H) The mRNA levels of CTNNB1 in NEAT1-knock down cells were detected using RT-qPCR. ****P<0.0001 vs. NC (n=3). (I) The mRNA levels of CTNNB1 in LDHB-knock down cells were detected using RT-qPCR. ***P<0.001 vs. NC (n=3). (J) The protein levels of β-catenin in NEAT1-knock down and LDHB-knock down cells were measured using western blotting (n=3). (K) Semi-quantitative analysis of the levels of β-catenin in NEAT1-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). (L) Semi-quantitative analysis of the levels of β-catenin in LDHB-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). NC, negative control; H3K18la, Histone H3K18 lactylation; si, small interfering (RNA); LDHB, lactate dehydrogenase B; CTNNB1, β-catenin gene; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Figure Legend Snippet: Knocking down long non-coding RNA NEAT1 inhibits cellular protein lactylation and downregulates β-catenin in hemangioma cells. (A) Cellular lactate levels were measured after NEAT1 knockdown; ****P<0.0001 vs. NC (n=3). (B) The level of pan-lactylation in NEAT1-knock down cells were detected by western blotting (n=3). (C) H3K18 lactylation in NEAT1-knock down cells was detected by western blotting (n=3). (D) Quantitative analysis of the levels of H3K18la in the western blotting results; *P<0.05, **P<0.01 vs. NC (n=3). (E) The knockdown efficiency of LDHB was detected by western blotting; *P<0.05 (n=3). (F) The viability of LDHB-knock down cells was detected via Cell Counting Kit-8 assay; **P<0.01 (n=3). (G) Colony formation ability of LDHB-knock down cells was detected via a colony formation assay; **P<0.01 (n=3). (H) The mRNA levels of CTNNB1 in NEAT1-knock down cells were detected using RT-qPCR. ****P<0.0001 vs. NC (n=3). (I) The mRNA levels of CTNNB1 in LDHB-knock down cells were detected using RT-qPCR. ***P<0.001 vs. NC (n=3). (J) The protein levels of β-catenin in NEAT1-knock down and LDHB-knock down cells were measured using western blotting (n=3). (K) Semi-quantitative analysis of the levels of β-catenin in NEAT1-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). (L) Semi-quantitative analysis of the levels of β-catenin in LDHB-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). NC, negative control; H3K18la, Histone H3K18 lactylation; si, small interfering (RNA); LDHB, lactate dehydrogenase B; CTNNB1, β-catenin gene; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Techniques Used: Knockdown, Western Blot, Cell Counting, Colony Assay, Quantitative RT-PCR, Negative Control, Small Interfering RNA, Reverse Transcription, Real-time Polymerase Chain Reaction

Long non-coding RNA NEAT1 promotes the H3K18 lactylation of the CTNNB1 promoter and the blockade of lactylation downregulates β-catenin in hemangioma cells. H3K18 lactylation levels of the CTNNB1 promoter in (A) 2-DG- or oxamate-treated cells and (B) lactate-treated, (C) NEAT1-knock down and (D) LDHB-knock down hemangioma cells were evaluated using ChIP-PCR; *P<0.05, **P<0.01, ***P<0.001 vs. NC (n=3). The CTNNB1 mRNA levels in (E) 2-DG- and oxamate-treated cells as well as (F) lactate-treated cells were detected by RT-qPCR; **P<0.01, ****P<0.0001 vs. NC (n=3). The protein levels of β-catenin in (G) 2-DG- and oxamate-treated cells were detected by western blotting (n=3). (H) Semi-quantitative analysis of the levels of β-catenin in 2-DG and oxamate-treated cells detected by western blotting; ***P<0.001 vs. NC (n=3). (I) The protein levels of β-catenin in lactate-treated cells were detected by western blotting (n=3). (J) Semi-quantitative analysis of the levels of β-catenin in lactate-treated cells detected by western blotting; *P<0.05 vs. NC (n=3). 2-DG, 2-Deoxy-D-glucose; NC, negative control; CTNNB1, β-catenin gene; LDHB, lactate dehydrogenase B; ChIP, chromatin immunoprecipitation; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; si, small interfering (RNA).

Figure Legend Snippet: Long non-coding RNA NEAT1 promotes the H3K18 lactylation of the CTNNB1 promoter and the blockade of lactylation downregulates β-catenin in hemangioma cells. H3K18 lactylation levels of the CTNNB1 promoter in (A) 2-DG- or oxamate-treated cells and (B) lactate-treated, (C) NEAT1-knock down and (D) LDHB-knock down hemangioma cells were evaluated using ChIP-PCR; *P<0.05, **P<0.01, ***P<0.001 vs. NC (n=3). The CTNNB1 mRNA levels in (E) 2-DG- and oxamate-treated cells as well as (F) lactate-treated cells were detected by RT-qPCR; **P<0.01, ****P<0.0001 vs. NC (n=3). The protein levels of β-catenin in (G) 2-DG- and oxamate-treated cells were detected by western blotting (n=3). (H) Semi-quantitative analysis of the levels of β-catenin in 2-DG and oxamate-treated cells detected by western blotting; ***P<0.001 vs. NC (n=3). (I) The protein levels of β-catenin in lactate-treated cells were detected by western blotting (n=3). (J) Semi-quantitative analysis of the levels of β-catenin in lactate-treated cells detected by western blotting; *P<0.05 vs. NC (n=3). 2-DG, 2-Deoxy-D-glucose; NC, negative control; CTNNB1, β-catenin gene; LDHB, lactate dehydrogenase B; ChIP, chromatin immunoprecipitation; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; si, small interfering (RNA).

Techniques Used: Knockdown, Quantitative RT-PCR, Western Blot, Negative Control, Chromatin Immunoprecipitation, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

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.

Figure Legend 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.

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

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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: Knocking down long non-coding RNA NEAT1 inhibits cellular protein lactylation and downregulates β-catenin in hemangioma cells. (A) Cellular lactate levels were measured after NEAT1 knockdown; ****P<0.0001 vs. NC (n=3). (B) The level of pan-lactylation in NEAT1-knock down cells were detected by western blotting (n=3). (C) H3K18 lactylation in NEAT1-knock down cells was detected by western blotting (n=3). (D) Quantitative analysis of the levels of H3K18la in the western blotting results; *P<0.05, **P<0.01 vs. NC (n=3). (E) The knockdown efficiency of LDHB was detected by western blotting; *P<0.05 (n=3). (F) The viability of LDHB-knock down cells was detected via Cell Counting Kit-8 assay; **P<0.01 (n=3). (G) Colony formation ability of LDHB-knock down cells was detected via a colony formation assay; **P<0.01 (n=3). (H) The mRNA levels of CTNNB1 in NEAT1-knock down cells were detected using RT-qPCR. ****P<0.0001 vs. NC (n=3). (I) The mRNA levels of CTNNB1 in LDHB-knock down cells were detected using RT-qPCR. ***P<0.001 vs. NC (n=3). (J) The protein levels of β-catenin in NEAT1-knock down and LDHB-knock down cells were measured using western blotting (n=3). (K) Semi-quantitative analysis of the levels of β-catenin in NEAT1-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). (L) Semi-quantitative analysis of the levels of β-catenin in LDHB-knock down cells detected by western blotting; ***P<0.001 vs. NC (n=3). NC, negative control; H3K18la, Histone H3K18 lactylation; si, small interfering (RNA); LDHB, lactate dehydrogenase B; CTNNB1, β-catenin gene; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

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, Western Blot, Cell Counting, Colony Assay, Quantitative RT-PCR, Negative Control, Small Interfering RNA, Reverse Transcription, Real-time Polymerase Chain Reaction

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 promotes the H3K18 lactylation of the CTNNB1 promoter and the blockade of lactylation downregulates β-catenin in hemangioma cells. H3K18 lactylation levels of the CTNNB1 promoter in (A) 2-DG- or oxamate-treated cells and (B) lactate-treated, (C) NEAT1-knock down and (D) LDHB-knock down hemangioma cells were evaluated using ChIP-PCR; *P<0.05, **P<0.01, ***P<0.001 vs. NC (n=3). The CTNNB1 mRNA levels in (E) 2-DG- and oxamate-treated cells as well as (F) lactate-treated cells were detected by RT-qPCR; **P<0.01, ****P<0.0001 vs. NC (n=3). The protein levels of β-catenin in (G) 2-DG- and oxamate-treated cells were detected by western blotting (n=3). (H) Semi-quantitative analysis of the levels of β-catenin in 2-DG and oxamate-treated cells detected by western blotting; ***P<0.001 vs. NC (n=3). (I) The protein levels of β-catenin in lactate-treated cells were detected by western blotting (n=3). (J) Semi-quantitative analysis of the levels of β-catenin in lactate-treated cells detected by western blotting; *P<0.05 vs. NC (n=3). 2-DG, 2-Deoxy-D-glucose; NC, negative control; CTNNB1, β-catenin gene; LDHB, lactate dehydrogenase B; ChIP, chromatin immunoprecipitation; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; si, small interfering (RNA).

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Negative Control, Chromatin Immunoprecipitation, Reverse Transcription, Real-time Polymerase Chain Reaction, 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.

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

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