Journal: Journal of Thoracic Disease

Article Title: 4-phenylbutyric acid re-trafficking hERG/G572R channel protein by modulating the endoplasmic reticulum stress-associated chaperones and endoplasmic reticulum-associated degradation gene

doi: 10.21037/jtd-23-1252

Figure Lengend Snippet: 4-PBA increased the current density of the hERG/G572R channel. (A) Representative whole-cell patch-clamp current traces were recorded using the top voltage protocol with and without 5 mM 4-PBA treatment. (B) Current density-voltage relationship of tail current density (pA/pF) in WT/G572R and WT/G572R + 4-PBA drug treatment groups. (C) Statistical graph of tail current at 0, 10, and 20 mV. Each scatter represents independent cell data. WT/G572R, n=5; WT/G572R + 4-PBA, n=5. **, P<0.01; ***, P<0.001; ****, P<0.0001. WT, wild type; 4-PBA, 4-phenylbutyric acid; hERG, human ether-à-go-go-related gene.

Article Snippet: 4-PBA (cat. no. P831259) was purchased from Macklin (Shanghai, China).

Techniques: Patch Clamp

Journal: Journal of Thoracic Disease

Article Title: 4-phenylbutyric acid re-trafficking hERG/G572R channel protein by modulating the endoplasmic reticulum stress-associated chaperones and endoplasmic reticulum-associated degradation gene

doi: 10.21037/jtd-23-1252

Figure Lengend Snippet: 4-PBA upregulated the expression of the hERG/G572R channel. (A) WB results showing protein expression of WT, G572R, and WT/G572R transfected cells, and protein expression of G572R and WT/G572R transfected cells after 4-PBA treatment. (B) Optical density analysis of the mature (155 kDa) form of hERG protein in 4-PBA-untreated control and 4-PBA-treated experimental groups. (C) hERG qPCR mRNA expression in 4-PBA-untreated control and 4-PBA-treated experimental groups. ns, non-significant; *, P<0.05; **, P<0.01; ****, P<0.0001. hERG, human ether-à-go-go-related gene; 4-PBA, 4-phenylbutyric acid; WT, wild type; WB, western blot; qPCR, quantitative real-time polymerase chain reaction; mRNA, messenger RNA.

Article Snippet: 4-PBA (cat. no. P831259) was purchased from Macklin (Shanghai, China).

Techniques: Expressing, Transfection, Western Blot, Real-time Polymerase Chain Reaction

Journal: Journal of Thoracic Disease

Article Title: 4-phenylbutyric acid re-trafficking hERG/G572R channel protein by modulating the endoplasmic reticulum stress-associated chaperones and endoplasmic reticulum-associated degradation gene

doi: 10.21037/jtd-23-1252

Figure Lengend Snippet: 4-PBA inhibited the expression of ATF6, GRP78, GRP94, and CRT/CNX. (A) WB results showing protein expression of ATF6, GRP78, and GRP94 in WT, WT/G572R, and WT/G572R + 5 mM 4-PBA cell lines. (B) Statistical analysis of the WB protein optical density of ATF6 in WT, WT/G572R, WT/G572R + 5 mM 4-PBA. (C) Statistical analysis of the WB protein optical density of GRP78 in WT, WT/G572R, and WT/G572R + 5 mM 4-PBA. (D) Statistical analysis of the WB protein optical density of GRP94 in WT, WT/G572R, and WT/G572R + 5 mM 4-PBA. (E) ATF6 mRNA expression levels in WT, WT/G572R untreated control, and WT/G572R + 4-PBA experimental groups. (F) GRP78 mRNA expression levels in WT, WT/G572R untreated control, and WT/G572R + 4-PBA experimental groups. (G) GRP94 mRNA expression levels in WT, WT/G572R untreated control, and WT/G572R + 4-PBA experimental groups. (H) CRT mRNA expression levels in WT, WT/G572R untreated control, and WT/G572R + 4-PBA experimental groups. (I) CNX mRNA expression levels in WT, WT/G572R untreated control, and WT/G572R + 4-PBA experimental groups. ns, non-significant; *, P<0.05; **, P<0.01; ****, P<0.0001. hERG, human ether-à-go-go-related gene; 4-PBA, 4-phenylbutyric acid; ATF6, activating transcription factor 6; WT, wild type; CRT, calreticulin; CNX, calnexin; WB, western blot; mRNA, messenger RNA.

Article Snippet: 4-PBA (cat. no. P831259) was purchased from Macklin (Shanghai, China).

Techniques: Expressing, Western Blot

Journal: Journal of Thoracic Disease

Article Title: 4-phenylbutyric acid re-trafficking hERG/G572R channel protein by modulating the endoplasmic reticulum stress-associated chaperones and endoplasmic reticulum-associated degradation gene

doi: 10.21037/jtd-23-1252

Figure Lengend Snippet: 4-PBA inhibited ERAD gene HRD1. (A) WB results showing HRD1 protein expression in WT, WT/G572R, and WT/G572R + 5 mM 4-PBA cell lines. (B) Statistical graph of HRD1 protein expression in WT, WT/G572R, and WT/G572R + 5 mM 4-PBA cell lines. (C) Statistical analysis of HRD1 mRNA expression in WT, WT/G572R and WT/G572R + 5 mM 4-PBA cell lines. (D) co-IP of GRP78, GRP94, HRD1, and hERG (WT/G572R)-FLAG channel. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. hERG, human ether-à-go-go-related gene; 4-PBA, 4-phenylbutyric acid; HRD1, 3-hydroxy-3-methylglutaryl coenzyme A reductase degradation protein 1; WT, wild type; WB, western blot; IP, immunoprecipitation; co-IP, co-immunoprecipitation; mRNA, messenger RNA.

Article Snippet: 4-PBA (cat. no. P831259) was purchased from Macklin (Shanghai, China).

Techniques: Expressing, Co-Immunoprecipitation Assay, Western Blot, Immunoprecipitation

Journal: Cell & Bioscience

Article Title: Blocking the interaction between interleukin-17A and endoplasmic reticulum stress in macrophage attenuates retinal neovascularization in oxygen-induced retinopathy

doi: 10.1186/s13578-021-00593-6

Figure Lengend Snippet: ER stress promoted production of IL-17A in macrophages exposed to hypoxia both in vivo and in vitro. a, b Effects of different concentrations of TM (0, 0.1, 0.5, 1.0 μg/μl) on GRP78, ATF-4, CHOP, IL-17A mRNA ( a ) and protein ( b ) expression levels in normal retinas. c, d Effects of different concentrations of 4-PBA (0, 0.1, 1, 10 nmol/μl) on GRP78, ATF-4, IL-17A mRNA ( c ) and protein ( d ) expression levels in OIR retinas. e Effects of different concentrations of TM (0, 0.1, 1, 2, 4, 6 μg/ml) on GRP78, ATF-4 and IL-17A protein expression levels in macrophages cultured under normoxic conditions. f, g Effects of different concentrations of 4-PBA (0, 0.1, 1, 5, 10 nmol/ml) on GRP78, ATF-4, CHOP, IL-17A mRNA ( f ) and protein ( g ) expression levels in macrophages cultured under hypoxic conditions. β-actin served as an endogenous reference for normalization. Data are shown as mean ± SEM, n = 6–8 per group for Real-time RT-PCR, n = 3 per group for western blotting. Each experiment repeated three times. ns, no significance. * P < 0.05 and **P < 0.01 compared with control groups

Article Snippet: Dimethylsulfoxide (DMSO) (Cat: D2650), 4-PBA (Cat: P21005) was purchased from Sigma (St.Louis, MO, USA).

Techniques: In Vivo, In Vitro, Expressing, Cell Culture, Quantitative RT-PCR, Western Blot

Journal: Cell & Bioscience

Article Title: Blocking the interaction between interleukin-17A and endoplasmic reticulum stress in macrophage attenuates retinal neovascularization in oxygen-induced retinopathy

doi: 10.1186/s13578-021-00593-6

Figure Lengend Snippet: Effects of IL-17ANab or 4-PBA on ER stress markers and IL-17A production in OIR retinas. a Western blotting analysis of GRP78, ATF-4, CHOP and IL-17A protein levels in normal retinas and OIR retinas pretreated with 4-PBA (10 nmol/μl) or IL-17ANab (1.0 μg/μl) or vehicle. b–d Immunofluorescent staining of F4/80 (red) and ATF-4 ( b , green), GRP78 ( c , green), IL-17A (d, green) in OIR retinas treated with 4-PBA or IL-17NAab or vehicle. Boxed areas are magnified in the images. Yellow represents co-localization of ATF-4 or GRP78 and macrophages infltration. Scale bars, 100 μm. β-actin served as an endogenous reference for normalization. Data are shown as mean ± SEM, n = 6–8 per group for Real-time RT-PCR, n = 3 per group for western blotting. Each experiment repeated three times. * P < 0.05 and **P < 0.01 compared with control groups or each other. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer

Article Snippet: Dimethylsulfoxide (DMSO) (Cat: D2650), 4-PBA (Cat: P21005) was purchased from Sigma (St.Louis, MO, USA).

Techniques: Western Blot, Staining, Quantitative RT-PCR

Journal: Cell & Bioscience

Article Title: Blocking the interaction between interleukin-17A and endoplasmic reticulum stress in macrophage attenuates retinal neovascularization in oxygen-induced retinopathy

doi: 10.1186/s13578-021-00593-6

Figure Lengend Snippet: Inhibition of ER stress or IL-17A alleviated RNV. a Immunofluorescence staining of fluorescein-conjugated lectin in retinal flat-mounts of normal and OIR WT mice at P17 treated with 4-PBA or IL-17ANab or vehicle, as well as in retinal flat-mounts of normal and OIR IL-17A KO mice. Areas of vaso-obliteration (within yellow lines) and NV (red) were quantified. b, c Quantification of the avascular area ( b ) and NV area ( c ) of retinal flat mounts. N = 6 per group. ns, no significance. ***P < 0.001 compared with OIR (WT)-vehicle group. ### P < 0.001 compared with OIR (WT)-control group. Scale bars, 500 μm. Data are shown as mean ± SEM. ns, no significance

Article Snippet: Dimethylsulfoxide (DMSO) (Cat: D2650), 4-PBA (Cat: P21005) was purchased from Sigma (St.Louis, MO, USA).

Techniques: Inhibition, Immunofluorescence, Staining

Journal: Cell & Bioscience

Article Title: Blocking the interaction between interleukin-17A and endoplasmic reticulum stress in macrophage attenuates retinal neovascularization in oxygen-induced retinopathy

doi: 10.1186/s13578-021-00593-6

Figure Lengend Snippet: ER stress promoted the activation of TXNIP/NLRP3 pathway in vivo and in vitro. a, b Effects of TM (0.5 μg/μl) on TXNIP, NLRP3 and IL-1β mRNA ( a ) and protein ( b ) expression levels in normal retinas. c, d Alterations of TXNIP, NLRP3 and IL-1β mRNA ( c ) and protein ( d ) levels in normal and OIR retinas pretreated with 4-PBA (10 nmol/μl) or vehicle. e, f Effects of TM (4 μg/ml) on TXNIP, NLRP3 and IL-1β mRNA ( e ) and protein ( f ) expression levels in normal macrophages. g, h Changes of TXNIP, NLRP3 and IL-1β mRNA ( g ) and protein ( h ) expression levels in macrophages cultured under normoxic or hypoxic conditions pretreated with 4-PBA (10 nmol/ml) or vehicle. β-actin served as an endogenous reference for normalization. Data are shown as mean ± SEM, n = 6–8 per group for Real-time RT-PCR, n = 3 per group for western blotting. Each experiment repeated three times. ns, no significance. *P < 0.05 and **P < 0.01 compared with control groups or each other

Article Snippet: Dimethylsulfoxide (DMSO) (Cat: D2650), 4-PBA (Cat: P21005) was purchased from Sigma (St.Louis, MO, USA).

Techniques: Activation Assay, In Vivo, In Vitro, Expressing, Cell Culture, Quantitative RT-PCR, Western Blot