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(A) UMAP visualization of major cell clusters identified from single cell RNA sequencing (scRNA-seq) of NT2.5-LM breast-to-lung metastatic tumors after 3 weeks of treatment with: vehicle (V), Entinostat (E), anti-CTLA-4 + anti-PD-1 immune checkpoint inhibitors (PC), and Entinostat + anti-CTLA-4 + anti-PD-1 combination (EPC). Subclusters of MDSCs identified: G-MDSCs and M-MDSCs. (B) Iterative logistic regression analyses conducted on the major MDSC cluster comparing the Vehicle (V) and Entinostat + ICIs combination treatment (EPC). (C) Expression of Malat1 / MALAT1 in G- and M-MDSCs isolated from lung metastases of NT2.5-LM mice treated with vehicle vs. Entinostat for 3 weeks (left, n=2 from 5 pooled mice per treatment group), J774M murine MDSC-like cell line treated with Entinostat for 24 hours (middle, n=3), and human PBMC-derived MDSCs (hMDSCs) treated with Entinostat for 24 hours (right, n=3). (D) Median Fluorescence Intensity expression of Malat1 in CD45 + CD11b + MHC-II - F4/80 - Ly6G - Ly6C hi cells from lung metastases in 4T1 mice, treated with vehicle (V), Entinostat (E), and Entinostat + ICIs (EPC) for 3 weeks. (E) Expression of Malat1 in J774M cell line treated with various concentrations of various HDAC inhibitors, with corresponding targeted class and specific HDACs. One-way ANOVA for (C, E: all statistically significant with p<0.05, unless indicated as non-significant (ns)), Kruskal-Wallis test with Dunn’s correction for (D). * p<0.05, *** p< 0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = <t>RGFP966;</t> TMP = TMP269; SIS = SIS17.

Rgfp966, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 95/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells"

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

Journal: bioRxiv

doi: 10.64898/2026.03.23.713743

Figure Legend Snippet: (A) UMAP visualization of major cell clusters identified from single cell RNA sequencing (scRNA-seq) of NT2.5-LM breast-to-lung metastatic tumors after 3 weeks of treatment with: vehicle (V), Entinostat (E), anti-CTLA-4 + anti-PD-1 immune checkpoint inhibitors (PC), and Entinostat + anti-CTLA-4 + anti-PD-1 combination (EPC). Subclusters of MDSCs identified: G-MDSCs and M-MDSCs. (B) Iterative logistic regression analyses conducted on the major MDSC cluster comparing the Vehicle (V) and Entinostat + ICIs combination treatment (EPC). (C) Expression of Malat1 / MALAT1 in G- and M-MDSCs isolated from lung metastases of NT2.5-LM mice treated with vehicle vs. Entinostat for 3 weeks (left, n=2 from 5 pooled mice per treatment group), J774M murine MDSC-like cell line treated with Entinostat for 24 hours (middle, n=3), and human PBMC-derived MDSCs (hMDSCs) treated with Entinostat for 24 hours (right, n=3). (D) Median Fluorescence Intensity expression of Malat1 in CD45 + CD11b + MHC-II - F4/80 - Ly6G - Ly6C hi cells from lung metastases in 4T1 mice, treated with vehicle (V), Entinostat (E), and Entinostat + ICIs (EPC) for 3 weeks. (E) Expression of Malat1 in J774M cell line treated with various concentrations of various HDAC inhibitors, with corresponding targeted class and specific HDACs. One-way ANOVA for (C, E: all statistically significant with p<0.05, unless indicated as non-significant (ns)), Kruskal-Wallis test with Dunn’s correction for (D). * p<0.05, *** p< 0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Techniques Used: Single Cell, RNA Sequencing, Expressing, Isolation, Derivative Assay, Fluorescence

(A) Western blots of Tyr705 phosphorylated-STAT3 (pSTAT3), STAT3, and Beta-actin in G-MDSCs isolated from NT2.5-LM metastatic lung tumors. G-MDSCs were isolated with Ly-6G+ magnetic beads from single cell suspensions of dissociated metastatic lung tumors and stimulated with LPS (2 hours, 1 ug/mL). NT2.5-LM tumor-bearing mice were treated with vehicle vs. Entinostat (5 mg/kg, 5x/wk) for 3 weeks. Each lane represents one mouse (n=5 per treatment group). 50 ug of total protein lysate loaded in each lane. Band density quantified with Image J and adjusted density normalized to Beta-actin (right), outlier mouse #2 in Vehicle removed with ROUT at threshold Q=1%. Two-way ANOVA, * p<0.05. (B) Western blot of pSTAT3, STAT3, and Beta-actin in J774M cells after 24 hours of Entinostat treatment and IFN-γ stimulation (30 min, 20 ng/mL). 20 ug of total protein lysate loaded in each lane. (C) Western blot of pSTAT3, STAT3, and Beta-actin in hMDSCs after 24 hours of Entinostat treatment. 50 ug of total protein lysate loaded in each lane. (D) Representative flow plots (left) and quantification (right, n=3) of J774M STAT3-responsive GFP reporter after 18 hours of pre-treatment with various concentrations of various HDAC inhibitors and IFN-γ stimulation for 5-7 hours (20 ng/mL), indicative of percentage of STAT3 activation. One-way ANOVA for (D: all statistically significant with p<0.05, unless indicated as non-significant (ns)). ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Figure Legend Snippet: (A) Western blots of Tyr705 phosphorylated-STAT3 (pSTAT3), STAT3, and Beta-actin in G-MDSCs isolated from NT2.5-LM metastatic lung tumors. G-MDSCs were isolated with Ly-6G+ magnetic beads from single cell suspensions of dissociated metastatic lung tumors and stimulated with LPS (2 hours, 1 ug/mL). NT2.5-LM tumor-bearing mice were treated with vehicle vs. Entinostat (5 mg/kg, 5x/wk) for 3 weeks. Each lane represents one mouse (n=5 per treatment group). 50 ug of total protein lysate loaded in each lane. Band density quantified with Image J and adjusted density normalized to Beta-actin (right), outlier mouse #2 in Vehicle removed with ROUT at threshold Q=1%. Two-way ANOVA, * p<0.05. (B) Western blot of pSTAT3, STAT3, and Beta-actin in J774M cells after 24 hours of Entinostat treatment and IFN-γ stimulation (30 min, 20 ng/mL). 20 ug of total protein lysate loaded in each lane. (C) Western blot of pSTAT3, STAT3, and Beta-actin in hMDSCs after 24 hours of Entinostat treatment. 50 ug of total protein lysate loaded in each lane. (D) Representative flow plots (left) and quantification (right, n=3) of J774M STAT3-responsive GFP reporter after 18 hours of pre-treatment with various concentrations of various HDAC inhibitors and IFN-γ stimulation for 5-7 hours (20 ng/mL), indicative of percentage of STAT3 activation. One-way ANOVA for (D: all statistically significant with p<0.05, unless indicated as non-significant (ns)). ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Techniques Used: Western Blot, Isolation, Magnetic Beads, Single Cell, Activation Assay

(A) Representative brightfield images of J774M cells after 72 hours of Entinostat and Stattic treatment. Scale bars = 300 um. (B) Representative flow cytometry plots and (C) percentage of apoptotic (Annexin V+, Propidium Iodide -) J774M cells after 72 hours of Entinostat and Stattic treatment (n=3). (D) Representative flow cytometry plots and (E) percentage of apoptotic hMDSCs after 48 hours of Entinostat and Stattic treatment (n=3). (F) Expression of anti-apoptotic genes ( Bcl2a1a, Bcl2a1b, Bcl2l1 ; green) and pro-apoptotic genes ( Bcl6, Bid, Bax ; red) in J774M cells after 24 hours of Entinostat treatment (n=3). (G) Western blot of pro-apoptotic BCL-xL, anti-apoptotic BID, and Beta-actin in J774M cells after 24 hours of Entinostat treatment and IFN-γ stimulation (30 min, 20 ng/mL). 20 ug of total protein lysate loaded in each lane. (H) Representative phase + NucView488 (top) and NucView488 only (bottom) live cell images at 20x magnification of J774M cells after 48 hours of various HDAC inhibitor treatments, indicative of active caspase 3/7. Scale bars = 200 um. (I) Total Integrated Intensity (TII) live cell imaging timecourse quantification of NucView488 fluorescence in J774M cells after 48 hours of various concentrations of Entinostat treatment and (J) various HDAC inhibitor treatments, indicative of active caspase 3/7 (n=3), with TII calculated from 9 images for each replicate. Statistical significance indicated for 48-hour timepoint by One-way ANOVA. ns = not significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; CHL = Chlopynostat; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269.

Figure Legend Snippet: (A) Representative brightfield images of J774M cells after 72 hours of Entinostat and Stattic treatment. Scale bars = 300 um. (B) Representative flow cytometry plots and (C) percentage of apoptotic (Annexin V+, Propidium Iodide -) J774M cells after 72 hours of Entinostat and Stattic treatment (n=3). (D) Representative flow cytometry plots and (E) percentage of apoptotic hMDSCs after 48 hours of Entinostat and Stattic treatment (n=3). (F) Expression of anti-apoptotic genes ( Bcl2a1a, Bcl2a1b, Bcl2l1 ; green) and pro-apoptotic genes ( Bcl6, Bid, Bax ; red) in J774M cells after 24 hours of Entinostat treatment (n=3). (G) Western blot of pro-apoptotic BCL-xL, anti-apoptotic BID, and Beta-actin in J774M cells after 24 hours of Entinostat treatment and IFN-γ stimulation (30 min, 20 ng/mL). 20 ug of total protein lysate loaded in each lane. (H) Representative phase + NucView488 (top) and NucView488 only (bottom) live cell images at 20x magnification of J774M cells after 48 hours of various HDAC inhibitor treatments, indicative of active caspase 3/7. Scale bars = 200 um. (I) Total Integrated Intensity (TII) live cell imaging timecourse quantification of NucView488 fluorescence in J774M cells after 48 hours of various concentrations of Entinostat treatment and (J) various HDAC inhibitor treatments, indicative of active caspase 3/7 (n=3), with TII calculated from 9 images for each replicate. Statistical significance indicated for 48-hour timepoint by One-way ANOVA. ns = not significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; CHL = Chlopynostat; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269.

Techniques Used: Flow Cytometry, Expressing, Western Blot, Live Cell Imaging, Fluorescence

Control:

Article Title: HOX code-based stratification reveals RUNX1T1-HDAC reprogramming as a targetable driver of lineage plasticity across cancers.
Article Snippet: Cancer remains a leading cause of death worldwide, with lineage plasticity emerging as a hallmark that drives therapy resistance and tumor progression by enabling cancer cells to alter identity and evade targeted therapies.. Although genomic and transcriptomic aberrations correlate with lineage plasticity, the absence of scalable crosscancer markers to rapidly identify plastic subtypes has limited predictive utility.. Homeobox (HOX) genes encode transcription factors that define tissue identity through distinct expression patterns, or HOX codes, within specific lineages.

In Vitro:

Article Title: DNTTIP1 drives leukaemogenesis through MiDAC‐mediated epigenetic silencing of BMF
Article Snippet: .. Drugs for in vitro studies were stored at ‒20°C at 10 mM in Dimethyl sulfoxide (DMSO) and listed as follows: cytarabine (Ara‐C) (TargetMol, T1272), daunorubicin hydrochloride (DNR) (TargetMol, T1511), doxorubicin (DOX) (TargetMol, T1456), vincristine sulphate (VCR) (TargetMol, T1270), venetoclax (ABT‐199) (Selleckchem, S8048), entinostat (MS‐275) (Selleckchem, S1053), tacedinaline (CI‐994) (Selleckchem, S2818), 5‐azacytidine (5‐AzaC) (MCE, HY‐10586), Olaparib (TargetMol, T3015), Merck60 (BRD‐6929) (MCE, HY‐100719), RGFP966 (MCE, HY‐13909) and FK228 (Romidepsin) (MCE, HY‐15149). ..

other:

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells
Article Snippet: Chemical inhibitors used in this study: Entinostat (Syndax Pharmaceuticals), Panobinostat (MedChemExpress, cat. #HY-10224), Belinostat (MedChemExpress, cat. #HY-10225), Vorinostat (MedChemExpress, cat. #HY-10221), Trichostatin A (MedChemExpress, cat. #HY-15144), Domatinostat (MedChemExpress, cat. #HY-16012A), Chlopynostat (MedChemExpress, cat. #HY-161464), Pyroxamide (MedChemExpress, cat. #HY-13216), Santacruzamate A (MedChemExpress, cat. #HY-N0931), RGFP966 (MedChemExpress, cat. #HY-13909), TMP269 (MedChemExpress, cat. #HY-18360), SIS17 (MedChemExpress, cat. #HY-128918), Stattic (MedChemExpress, cat. #HY-13818), Abemaciclib (MedChemExpress, cat. #HY-16297A), Palbociclib (MedChemExpress, cat. #50767).

Image Search Results

Journal: bioRxiv

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

doi: 10.64898/2026.03.23.713743

Figure Lengend Snippet: (A) UMAP visualization of major cell clusters identified from single cell RNA sequencing (scRNA-seq) of NT2.5-LM breast-to-lung metastatic tumors after 3 weeks of treatment with: vehicle (V), Entinostat (E), anti-CTLA-4 + anti-PD-1 immune checkpoint inhibitors (PC), and Entinostat + anti-CTLA-4 + anti-PD-1 combination (EPC). Subclusters of MDSCs identified: G-MDSCs and M-MDSCs. (B) Iterative logistic regression analyses conducted on the major MDSC cluster comparing the Vehicle (V) and Entinostat + ICIs combination treatment (EPC). (C) Expression of Malat1 / MALAT1 in G- and M-MDSCs isolated from lung metastases of NT2.5-LM mice treated with vehicle vs. Entinostat for 3 weeks (left, n=2 from 5 pooled mice per treatment group), J774M murine MDSC-like cell line treated with Entinostat for 24 hours (middle, n=3), and human PBMC-derived MDSCs (hMDSCs) treated with Entinostat for 24 hours (right, n=3). (D) Median Fluorescence Intensity expression of Malat1 in CD45 + CD11b + MHC-II - F4/80 - Ly6G - Ly6C hi cells from lung metastases in 4T1 mice, treated with vehicle (V), Entinostat (E), and Entinostat + ICIs (EPC) for 3 weeks. (E) Expression of Malat1 in J774M cell line treated with various concentrations of various HDAC inhibitors, with corresponding targeted class and specific HDACs. One-way ANOVA for (C, E: all statistically significant with p<0.05, unless indicated as non-significant (ns)), Kruskal-Wallis test with Dunn’s correction for (D). * p<0.05, *** p< 0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Article Snippet: Chemical inhibitors used in this study: Entinostat (Syndax Pharmaceuticals), Panobinostat (MedChemExpress, cat. #HY-10224), Belinostat (MedChemExpress, cat. #HY-10225), Vorinostat (MedChemExpress, cat. #HY-10221), Trichostatin A (MedChemExpress, cat. #HY-15144), Domatinostat (MedChemExpress, cat. #HY-16012A), Chlopynostat (MedChemExpress, cat. #HY-161464), Pyroxamide (MedChemExpress, cat. #HY-13216), Santacruzamate A (MedChemExpress, cat. #HY-N0931), RGFP966 (MedChemExpress, cat. #HY-13909), TMP269 (MedChemExpress, cat. #HY-18360), SIS17 (MedChemExpress, cat. #HY-128918), Stattic (MedChemExpress, cat. #HY-13818), Abemaciclib (MedChemExpress, cat. #HY-16297A), Palbociclib (MedChemExpress, cat. #50767).

Techniques: Single Cell, RNA Sequencing, Expressing, Isolation, Derivative Assay, Fluorescence

Journal: bioRxiv

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

doi: 10.64898/2026.03.23.713743

Figure Lengend Snippet: (A) Western blots of Tyr705 phosphorylated-STAT3 (pSTAT3), STAT3, and Beta-actin in G-MDSCs isolated from NT2.5-LM metastatic lung tumors. G-MDSCs were isolated with Ly-6G+ magnetic beads from single cell suspensions of dissociated metastatic lung tumors and stimulated with LPS (2 hours, 1 ug/mL). NT2.5-LM tumor-bearing mice were treated with vehicle vs. Entinostat (5 mg/kg, 5x/wk) for 3 weeks. Each lane represents one mouse (n=5 per treatment group). 50 ug of total protein lysate loaded in each lane. Band density quantified with Image J and adjusted density normalized to Beta-actin (right), outlier mouse #2 in Vehicle removed with ROUT at threshold Q=1%. Two-way ANOVA, * p<0.05. (B) Western blot of pSTAT3, STAT3, and Beta-actin in J774M cells after 24 hours of Entinostat treatment and IFN-γ stimulation (30 min, 20 ng/mL). 20 ug of total protein lysate loaded in each lane. (C) Western blot of pSTAT3, STAT3, and Beta-actin in hMDSCs after 24 hours of Entinostat treatment. 50 ug of total protein lysate loaded in each lane. (D) Representative flow plots (left) and quantification (right, n=3) of J774M STAT3-responsive GFP reporter after 18 hours of pre-treatment with various concentrations of various HDAC inhibitors and IFN-γ stimulation for 5-7 hours (20 ng/mL), indicative of percentage of STAT3 activation. One-way ANOVA for (D: all statistically significant with p<0.05, unless indicated as non-significant (ns)). ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Techniques: Western Blot, Isolation, Magnetic Beads, Single Cell, Activation Assay

Journal: bioRxiv

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

doi: 10.64898/2026.03.23.713743

Figure Lengend Snippet: (A) Representative brightfield images of J774M cells after 72 hours of Entinostat and Stattic treatment. Scale bars = 300 um. (B) Representative flow cytometry plots and (C) percentage of apoptotic (Annexin V+, Propidium Iodide -) J774M cells after 72 hours of Entinostat and Stattic treatment (n=3). (D) Representative flow cytometry plots and (E) percentage of apoptotic hMDSCs after 48 hours of Entinostat and Stattic treatment (n=3). (F) Expression of anti-apoptotic genes ( Bcl2a1a, Bcl2a1b, Bcl2l1 ; green) and pro-apoptotic genes ( Bcl6, Bid, Bax ; red) in J774M cells after 24 hours of Entinostat treatment (n=3). (G) Western blot of pro-apoptotic BCL-xL, anti-apoptotic BID, and Beta-actin in J774M cells after 24 hours of Entinostat treatment and IFN-γ stimulation (30 min, 20 ng/mL). 20 ug of total protein lysate loaded in each lane. (H) Representative phase + NucView488 (top) and NucView488 only (bottom) live cell images at 20x magnification of J774M cells after 48 hours of various HDAC inhibitor treatments, indicative of active caspase 3/7. Scale bars = 200 um. (I) Total Integrated Intensity (TII) live cell imaging timecourse quantification of NucView488 fluorescence in J774M cells after 48 hours of various concentrations of Entinostat treatment and (J) various HDAC inhibitor treatments, indicative of active caspase 3/7 (n=3), with TII calculated from 9 images for each replicate. Statistical significance indicated for 48-hour timepoint by One-way ANOVA. ns = not significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; CHL = Chlopynostat; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269.

Techniques: Flow Cytometry, Expressing, Western Blot, Live Cell Imaging, Fluorescence

ALKBH5/lincRNA-p21/FGFR2 promotes autophagy in response to ethanol-induced liver injury. ( A ) HepG2 cells were treated with/without 10 μM SAHA, 10 μM NaB, or 5 μM RGFP966 for 24 h, the expression of lincRNA-p21 was evaluated by RT-PCR. n=3. ( B ) The potential m 6 A sites in lincRNA-p21 predicted using SRAMP. ( C ) m 6 A RNA methylation levels of HepG2 cells with/without alcoholic hepatocyte injury ( P >0.05). n=6. ( D ) When HepG2 cells were exposed to ethanol, the expressions of lincRNA-p21, ALKBH5 and METTL3 were evaluated. The expressions of lincRNA-p21 and ALKBH5 were all down-regulated when HepG2 cells were exposed to 100 mM ethanol ( P <0.01, P <0.05). ( E ) The location of lincRNA-p21 in HepG2 cells demonstrated by the lincRNA-p21 FISH Probe. Scale bars: 50 μm. ( F ) The expressions of lincRNA-p21 was upregulated by ALKBH5 overexpression. n=3. ( G ) LincRNA-p21 was significantly enriched in ALKBH5-bound RNA. n=3. ( H )The methylation level of the 1237 nt site in lincRNA-p21 was evaluated using MeRIP-PCR. n=3. ( I ) Schematic diagram of ALKBH5 mediating m 6 A demethylation and upregulation of lincRNA-p21. The red downward arrow indicates decreased expression. ( J ) The mRNA and protein expression of FGFR2 was evaluated by RT-PCR and Western blot. FGFR2 protein was up-regulated by the overexpression of lincRNA-p21 (FGFR2/β-actin: 0.54 vs 0.20, P =0.001). ( K ) Fluorescence images of the autophagosomes. Scale bars: 10 μm. ( L ) Cell viability was evaluated using a CCK8 assay. n=3. Data represent mean±s.e.m. Significance was determined by two-tailed Student’s t -test. * P <0.05, ** P <0.01, *** P <0.001.

Journal: International Journal of Nanomedicine

Article Title: LincRNA-p21: A Double-Edged Sword in Ethanol-Induced Liver Damage and Its Nanoparticle Solution

doi: 10.2147/IJN.S577455

Figure Lengend Snippet: ALKBH5/lincRNA-p21/FGFR2 promotes autophagy in response to ethanol-induced liver injury. ( A ) HepG2 cells were treated with/without 10 μM SAHA, 10 μM NaB, or 5 μM RGFP966 for 24 h, the expression of lincRNA-p21 was evaluated by RT-PCR. n=3. ( B ) The potential m 6 A sites in lincRNA-p21 predicted using SRAMP. ( C ) m 6 A RNA methylation levels of HepG2 cells with/without alcoholic hepatocyte injury ( P >0.05). n=6. ( D ) When HepG2 cells were exposed to ethanol, the expressions of lincRNA-p21, ALKBH5 and METTL3 were evaluated. The expressions of lincRNA-p21 and ALKBH5 were all down-regulated when HepG2 cells were exposed to 100 mM ethanol ( P <0.01, P <0.05). ( E ) The location of lincRNA-p21 in HepG2 cells demonstrated by the lincRNA-p21 FISH Probe. Scale bars: 50 μm. ( F ) The expressions of lincRNA-p21 was upregulated by ALKBH5 overexpression. n=3. ( G ) LincRNA-p21 was significantly enriched in ALKBH5-bound RNA. n=3. ( H )The methylation level of the 1237 nt site in lincRNA-p21 was evaluated using MeRIP-PCR. n=3. ( I ) Schematic diagram of ALKBH5 mediating m 6 A demethylation and upregulation of lincRNA-p21. The red downward arrow indicates decreased expression. ( J ) The mRNA and protein expression of FGFR2 was evaluated by RT-PCR and Western blot. FGFR2 protein was up-regulated by the overexpression of lincRNA-p21 (FGFR2/β-actin: 0.54 vs 0.20, P =0.001). ( K ) Fluorescence images of the autophagosomes. Scale bars: 10 μm. ( L ) Cell viability was evaluated using a CCK8 assay. n=3. Data represent mean±s.e.m. Significance was determined by two-tailed Student’s t -test. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The HDAC inhibitors Vorinostat (SAHA)(#HY-10221, MCE, USA), sodium butyrate (NaB)(#S1999, Selleck, USA), and HDAC3 inhibitor RGFP966 (#HY-13909, MCE, USA) were used to study the mechanism of lincRNA-p21 regulation.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Methylation, Over Expression, Western Blot, Fluorescence, CCK-8 Assay, Two Tailed Test

Review

rgfp966 (MedChemExpress)

Structured Review

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