Review

polr2h (Santa Cruz Biotechnology)

Santa Cruz Biotechnology is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Santa Cruz Biotechnology polr2h

GLS1 interacts with POLR2E or <t>POLR2H.</t> A) Schematic of immunoprecipitation using mass spectrometry (IP–MS) to identify proteins associated with GLS1 in LO2 cells. B) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes. β‐actin served as the loading control. C–F) CoIP analysis of the interaction of GAC–Myc and POLR2H‐HA, KGA‐Flag and POLR2H‐HA, GAC–Myc and POLR2E‐HA and KGA‐Flag and POLR2E‐HA in HEK293T cells. β‐actin served as the loading control. G) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes treated with ethanol (600 m m ) for 24 h. β‐actin served as the loading control. H,I) CoIP analysis of the interaction of GAC–Myc or KGA‐Flag with endogenous POLR2H, GAC–Myc or KGA‐Flag with endogenous POLR2E treated with ethanol (400 m m ) for 24 h in HEK293T cells. β‐actin served as the loading control.

Polr2h, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/polr2h/pmc12677690-238-30-32?v=Santa+Cruz+Biotechnology
Average 91 stars, based on 3 article reviews

polr2h - by Bioz Stars, 2026-08

91/100 stars

Images

1) Product Images from "GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets"

Article Title: GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets

Journal: Advanced Science

doi: 10.1002/advs.202502738

GLS1 interacts with POLR2E or POLR2H. A) Schematic of immunoprecipitation using mass spectrometry (IP–MS) to identify proteins associated with GLS1 in LO2 cells. B) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes. β‐actin served as the loading control. C–F) CoIP analysis of the interaction of GAC–Myc and POLR2H‐HA, KGA‐Flag and POLR2H‐HA, GAC–Myc and POLR2E‐HA and KGA‐Flag and POLR2E‐HA in HEK293T cells. β‐actin served as the loading control. G) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes treated with ethanol (600 m m ) for 24 h. β‐actin served as the loading control. H,I) CoIP analysis of the interaction of GAC–Myc or KGA‐Flag with endogenous POLR2H, GAC–Myc or KGA‐Flag with endogenous POLR2E treated with ethanol (400 m m ) for 24 h in HEK293T cells. β‐actin served as the loading control.

Figure Legend Snippet: GLS1 interacts with POLR2E or POLR2H. A) Schematic of immunoprecipitation using mass spectrometry (IP–MS) to identify proteins associated with GLS1 in LO2 cells. B) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes. β‐actin served as the loading control. C–F) CoIP analysis of the interaction of GAC–Myc and POLR2H‐HA, KGA‐Flag and POLR2H‐HA, GAC–Myc and POLR2E‐HA and KGA‐Flag and POLR2E‐HA in HEK293T cells. β‐actin served as the loading control. G) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes treated with ethanol (600 m m ) for 24 h. β‐actin served as the loading control. H,I) CoIP analysis of the interaction of GAC–Myc or KGA‐Flag with endogenous POLR2H, GAC–Myc or KGA‐Flag with endogenous POLR2E treated with ethanol (400 m m ) for 24 h in HEK293T cells. β‐actin served as the loading control.

Techniques Used: Immunoprecipitation, Mass Spectrometry, Protein-Protein interactions, Control

GLS1 interacts with POLR2H or POLR2E to reduce the activity of RNA pol II. A,B) The protein complex structure of GLS1, POLR2H and POLR2E performed on the GROMACS platform. The corresponding location of the putative binding motif on GLS1 and POLR2H protein, GLS1 and POLR2E protein. (C) The truncated GLS1 variants. D,E) CoIP analysis of the interaction of truncated GLS1 proteins and POLR2E‐HA or POLR2H‐HA in lysates of HEK293T cells. β‐actin served as the loading control. F) RNA pol II activity of AML12 cells expressing pGL3‐Basic or pGL3‐Promoter with KGA WT or ΔKGA‐3. Cells were serum starved overnight followed by ethanol (400 m m ) stimulation for 1 h. n = 10. G,H) The truncated POLR2E or POLR2H variants. I,J) CoIP analysis of the interaction of truncated POLR2E or POLR2H proteins and KGA‐Flag in lysates of HEK293T cells. β‐actin served as the loading control. K) RNA pol II activity of AML12 cells expressing pGL3‐Basic or pGL3‐Promoter with GLS1 and POLR2E WT or ΔPOLR2E‐3 or POLR2H WT or ΔPOLR2H‐1. Cells were serum starved overnight followed by ethanol (400 m m ) stimulation for 1 h. n = 7–10. Data in (F) and (K) are presented as the mean ± SEM, determined by one‐way ANOVA and Fisher's LSD test.

Figure Legend Snippet: GLS1 interacts with POLR2H or POLR2E to reduce the activity of RNA pol II. A,B) The protein complex structure of GLS1, POLR2H and POLR2E performed on the GROMACS platform. The corresponding location of the putative binding motif on GLS1 and POLR2H protein, GLS1 and POLR2E protein. (C) The truncated GLS1 variants. D,E) CoIP analysis of the interaction of truncated GLS1 proteins and POLR2E‐HA or POLR2H‐HA in lysates of HEK293T cells. β‐actin served as the loading control. F) RNA pol II activity of AML12 cells expressing pGL3‐Basic or pGL3‐Promoter with KGA WT or ΔKGA‐3. Cells were serum starved overnight followed by ethanol (400 m m ) stimulation for 1 h. n = 10. G,H) The truncated POLR2E or POLR2H variants. I,J) CoIP analysis of the interaction of truncated POLR2E or POLR2H proteins and KGA‐Flag in lysates of HEK293T cells. β‐actin served as the loading control. K) RNA pol II activity of AML12 cells expressing pGL3‐Basic or pGL3‐Promoter with GLS1 and POLR2E WT or ΔPOLR2E‐3 or POLR2H WT or ΔPOLR2H‐1. Cells were serum starved overnight followed by ethanol (400 m m ) stimulation for 1 h. n = 7–10. Data in (F) and (K) are presented as the mean ± SEM, determined by one‐way ANOVA and Fisher's LSD test.

Techniques Used: Activity Assay, Binding Assay, Control, Expressing

GLS1 reduces hepatic alcoholic steatosis through interacting with POLR2E or POLR2H. A) TG levels of the primary hepatocytes expressing with KGA WT or ΔKGA‐3. Cells were serum starved overnight followed by ethanol (600 m m ) stimulation for 24 h. The amount was normalized to the protein content. n = 5‐6. B) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (A); β‐actin serves as a loading control. C) TG levels of the primary hepatocytes expressing with GLS1 and POLR2E WT or ΔPOLR2E‐3 or POLR2H WT or ΔPOLR2H‐1. Cells were serum starved overnight followed by ethanol (600 m m ) stimulation for 24 h. The amount was normalized to the protein content. n = 4–6. D) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (C); β‐actin serves as a loading control. E) Schematic illustrating the groups and procedures of the mouse model. C57BL/6J mice were injected with AAV8‐TBG‐GLS1 with POLR2E WT or POLR2H WT or their truncated variants (ΔPOLR2E‐3 or ΔPOLR2H‐1) via the tail vein for additional 4‐week pair or ethanol feeding. n = 8–10 biologically independent mice per group. F) Change curves of body weight of mice. G) Liver TG levels of mice in the indicated group in (E); n = 6‐7. H) Representative H&E staining of liver sections in the indicated group in (E). Scale bars, 100 and 200 µm. I) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (E); β‐actin serves as a loading control. Data in (A), (C), (G) are presented as the mean ± SEM, determined by one‐way ANOVA and Fisher's LSD test.

Figure Legend Snippet: GLS1 reduces hepatic alcoholic steatosis through interacting with POLR2E or POLR2H. A) TG levels of the primary hepatocytes expressing with KGA WT or ΔKGA‐3. Cells were serum starved overnight followed by ethanol (600 m m ) stimulation for 24 h. The amount was normalized to the protein content. n = 5‐6. B) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (A); β‐actin serves as a loading control. C) TG levels of the primary hepatocytes expressing with GLS1 and POLR2E WT or ΔPOLR2E‐3 or POLR2H WT or ΔPOLR2H‐1. Cells were serum starved overnight followed by ethanol (600 m m ) stimulation for 24 h. The amount was normalized to the protein content. n = 4–6. D) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (C); β‐actin serves as a loading control. E) Schematic illustrating the groups and procedures of the mouse model. C57BL/6J mice were injected with AAV8‐TBG‐GLS1 with POLR2E WT or POLR2H WT or their truncated variants (ΔPOLR2E‐3 or ΔPOLR2H‐1) via the tail vein for additional 4‐week pair or ethanol feeding. n = 8–10 biologically independent mice per group. F) Change curves of body weight of mice. G) Liver TG levels of mice in the indicated group in (E); n = 6‐7. H) Representative H&E staining of liver sections in the indicated group in (E). Scale bars, 100 and 200 µm. I) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (E); β‐actin serves as a loading control. Data in (A), (C), (G) are presented as the mean ± SEM, determined by one‐way ANOVA and Fisher's LSD test.

Techniques Used: Expressing, Western Blot, Control, Injection, Staining

Schematic diagram of glutamine‐regulated GLS1 coordinates RNA polymerase II manipulates AFLD. High‐protein diet reduces alcoholic hepatic steatosis through glutamine stabilizing GLS1 to regulate RNA pol II activity by interacting with POLR2E or POLR2H.

Figure Legend Snippet: Schematic diagram of glutamine‐regulated GLS1 coordinates RNA polymerase II manipulates AFLD. High‐protein diet reduces alcoholic hepatic steatosis through glutamine stabilizing GLS1 to regulate RNA pol II activity by interacting with POLR2E or POLR2H.

Techniques Used: Activity Assay

Image Search Results

Journal: Advanced Science

Article Title: GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets

doi: 10.1002/advs.202502738

Figure Lengend Snippet: GLS1 interacts with POLR2E or POLR2H. A) Schematic of immunoprecipitation using mass spectrometry (IP–MS) to identify proteins associated with GLS1 in LO2 cells. B) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes. β‐actin served as the loading control. C–F) CoIP analysis of the interaction of GAC–Myc and POLR2H‐HA, KGA‐Flag and POLR2H‐HA, GAC–Myc and POLR2E‐HA and KGA‐Flag and POLR2E‐HA in HEK293T cells. β‐actin served as the loading control. G) CoIP analysis of the interaction of endogenous GLS1 with endogenous POLR2E or POLR2H in the primary hepatocytes treated with ethanol (600 m m ) for 24 h. β‐actin served as the loading control. H,I) CoIP analysis of the interaction of GAC–Myc or KGA‐Flag with endogenous POLR2H, GAC–Myc or KGA‐Flag with endogenous POLR2E treated with ethanol (400 m m ) for 24 h in HEK293T cells. β‐actin served as the loading control.

Article Snippet: For analysis of endogenous proteins, the primary hepatocytes lysed by cell lysis IP buffer were incubated overnight at 4 °C with GLS1 (12855‐1; Proteintech) or POLR2E (sc‐390902; santa cruz) or POLR2H (sc‐398512; santa cruz) antibody and protein A+G Agarose beads.

Techniques: Immunoprecipitation, Mass Spectrometry, Protein-Protein interactions, Control

Journal: Advanced Science

Article Title: GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets

doi: 10.1002/advs.202502738

Figure Lengend Snippet: GLS1 interacts with POLR2H or POLR2E to reduce the activity of RNA pol II. A,B) The protein complex structure of GLS1, POLR2H and POLR2E performed on the GROMACS platform. The corresponding location of the putative binding motif on GLS1 and POLR2H protein, GLS1 and POLR2E protein. (C) The truncated GLS1 variants. D,E) CoIP analysis of the interaction of truncated GLS1 proteins and POLR2E‐HA or POLR2H‐HA in lysates of HEK293T cells. β‐actin served as the loading control. F) RNA pol II activity of AML12 cells expressing pGL3‐Basic or pGL3‐Promoter with KGA WT or ΔKGA‐3. Cells were serum starved overnight followed by ethanol (400 m m ) stimulation for 1 h. n = 10. G,H) The truncated POLR2E or POLR2H variants. I,J) CoIP analysis of the interaction of truncated POLR2E or POLR2H proteins and KGA‐Flag in lysates of HEK293T cells. β‐actin served as the loading control. K) RNA pol II activity of AML12 cells expressing pGL3‐Basic or pGL3‐Promoter with GLS1 and POLR2E WT or ΔPOLR2E‐3 or POLR2H WT or ΔPOLR2H‐1. Cells were serum starved overnight followed by ethanol (400 m m ) stimulation for 1 h. n = 7–10. Data in (F) and (K) are presented as the mean ± SEM, determined by one‐way ANOVA and Fisher's LSD test.

Techniques: Activity Assay, Binding Assay, Control, Expressing

Journal: Advanced Science

Article Title: GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets

doi: 10.1002/advs.202502738

Figure Lengend Snippet: GLS1 reduces hepatic alcoholic steatosis through interacting with POLR2E or POLR2H. A) TG levels of the primary hepatocytes expressing with KGA WT or ΔKGA‐3. Cells were serum starved overnight followed by ethanol (600 m m ) stimulation for 24 h. The amount was normalized to the protein content. n = 5‐6. B) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (A); β‐actin serves as a loading control. C) TG levels of the primary hepatocytes expressing with GLS1 and POLR2E WT or ΔPOLR2E‐3 or POLR2H WT or ΔPOLR2H‐1. Cells were serum starved overnight followed by ethanol (600 m m ) stimulation for 24 h. The amount was normalized to the protein content. n = 4–6. D) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (C); β‐actin serves as a loading control. E) Schematic illustrating the groups and procedures of the mouse model. C57BL/6J mice were injected with AAV8‐TBG‐GLS1 with POLR2E WT or POLR2H WT or their truncated variants (ΔPOLR2E‐3 or ΔPOLR2H‐1) via the tail vein for additional 4‐week pair or ethanol feeding. n = 8–10 biologically independent mice per group. F) Change curves of body weight of mice. G) Liver TG levels of mice in the indicated group in (E); n = 6‐7. H) Representative H&E staining of liver sections in the indicated group in (E). Scale bars, 100 and 200 µm. I) Western blots of ACLY, ACC, FASN levels in the primary hepatocytes described in (E); β‐actin serves as a loading control. Data in (A), (C), (G) are presented as the mean ± SEM, determined by one‐way ANOVA and Fisher's LSD test.

Techniques: Expressing, Western Blot, Control, Injection, Staining

Journal: Advanced Science

Article Title: GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets

doi: 10.1002/advs.202502738

Figure Lengend Snippet: Schematic diagram of glutamine‐regulated GLS1 coordinates RNA polymerase II manipulates AFLD. High‐protein diet reduces alcoholic hepatic steatosis through glutamine stabilizing GLS1 to regulate RNA pol II activity by interacting with POLR2E or POLR2H.

Techniques: Activity Assay

(A) TRExRTA BCBL-1 cells bearing the indicated constructs were treated with 25 μM etoposide (Etop; DNA damage agent) for 24 h, 2 μg/mL doxycycline (Dox; viral reactivation) for 24h, or 100 μg/mL cycloheximide (Chx; translational inhibitor) for 6 h, or mock treated with drug diluent and analyzed by western blotting. γH2AX is induced during DNA damage response, KSHV SOX is a viral early protein and marker for infection, and p27 is a short-lived control for translation inhibition. Vinculin is a loading control. Black arrow indicates full length strep-tagged TFIIB and gray arrow indicated full-length endogenous TFIIB. (B) Empty or TFIIB-2xStrep expressing TRExRTA BCBL-1 cells were treated with either 100 ng/mL TRAIL ligand or mock treated with TRAIL dilution buffer for 6 h before western blotting. Caspase-3 cleavage is a control for induction of cell death. (C) Samples in (A) were probed for Caspase-3 and Caspase-8 activation by cleavage. (D) TRExRTA BCBL-1 cells expressing either TFIIB-2xStrep or mutant TFIIB(D207A)-2xStrep were treated with 100 ng/mL TRAIL ligand or mock for 8 h, followed by western blotting. Black arrow indicates cleavage product. (E) TRExRTA BCBL-1 cells expressing TFIIB-2xStrep or mutant TFIIB(D207A)-2xStrep were pretreated with 80 μM of either the caspase-3 inhibitor Z-VAD-DEVD-FMK, the pan-caspase inhibitor Z-VAD-OMe-FMK, or a negative control cysteine protease inhibitor Z-FA-FMK for 2 h. Cells were then treated with either 500 ng/mL TRAIL ligand or mock treated with TRAIL dilution buffer for 7 h followed by western blotting. Boosted strep signal has gamma contrast enhanced for visualization of the cleavage product and GAPDH probed with mouse antibody is a loading control. (F) TFIIB-2xStrep or TFIIB(D207A)-2xStrep expressing TRExRTA BCBL-1 cells were treated with 660 ng/mL TRAIL for 7 h and RNA extracted and sequenced with ribodepletion and ERCC spike-in control RNA for normalization. Volcano plot shows differentially expressed genes in cells containing TFIIB(D207A)-2xStrep relative to cells containing wild-type TFIIB-2xStrep. Genes in red are significantly differentially regulated in TRAIL-treated cells with described links to apoptosis (see Table S1 ), genes in yellow are differentially expressed in both TRAIL-treated and untreated cells, and in blue are genes with unknown function or no known link to apoptosis that are differentially expressed during TRAIL-treatment.

Journal: bioRxiv

Article Title: The RNA polymerase II general transcription factor TFIIB is a target for transcriptome control during cellular stress and viral infection

doi: 10.1101/2024.01.16.575933

Figure Lengend Snippet: (A) TRExRTA BCBL-1 cells bearing the indicated constructs were treated with 25 μM etoposide (Etop; DNA damage agent) for 24 h, 2 μg/mL doxycycline (Dox; viral reactivation) for 24h, or 100 μg/mL cycloheximide (Chx; translational inhibitor) for 6 h, or mock treated with drug diluent and analyzed by western blotting. γH2AX is induced during DNA damage response, KSHV SOX is a viral early protein and marker for infection, and p27 is a short-lived control for translation inhibition. Vinculin is a loading control. Black arrow indicates full length strep-tagged TFIIB and gray arrow indicated full-length endogenous TFIIB. (B) Empty or TFIIB-2xStrep expressing TRExRTA BCBL-1 cells were treated with either 100 ng/mL TRAIL ligand or mock treated with TRAIL dilution buffer for 6 h before western blotting. Caspase-3 cleavage is a control for induction of cell death. (C) Samples in (A) were probed for Caspase-3 and Caspase-8 activation by cleavage. (D) TRExRTA BCBL-1 cells expressing either TFIIB-2xStrep or mutant TFIIB(D207A)-2xStrep were treated with 100 ng/mL TRAIL ligand or mock for 8 h, followed by western blotting. Black arrow indicates cleavage product. (E) TRExRTA BCBL-1 cells expressing TFIIB-2xStrep or mutant TFIIB(D207A)-2xStrep were pretreated with 80 μM of either the caspase-3 inhibitor Z-VAD-DEVD-FMK, the pan-caspase inhibitor Z-VAD-OMe-FMK, or a negative control cysteine protease inhibitor Z-FA-FMK for 2 h. Cells were then treated with either 500 ng/mL TRAIL ligand or mock treated with TRAIL dilution buffer for 7 h followed by western blotting. Boosted strep signal has gamma contrast enhanced for visualization of the cleavage product and GAPDH probed with mouse antibody is a loading control. (F) TFIIB-2xStrep or TFIIB(D207A)-2xStrep expressing TRExRTA BCBL-1 cells were treated with 660 ng/mL TRAIL for 7 h and RNA extracted and sequenced with ribodepletion and ERCC spike-in control RNA for normalization. Volcano plot shows differentially expressed genes in cells containing TFIIB(D207A)-2xStrep relative to cells containing wild-type TFIIB-2xStrep. Genes in red are significantly differentially regulated in TRAIL-treated cells with described links to apoptosis (see Table S1 ), genes in yellow are differentially expressed in both TRAIL-treated and untreated cells, and in blue are genes with unknown function or no known link to apoptosis that are differentially expressed during TRAIL-treatment.

Article Snippet: Western blotting was performed with Tris-buffered saline and 0.2% Tween-20 (TBS-T) using PageRuler Prestained protein ladder (10-180 kDa formulation; ThermoFisher) and the following primary and secondary antibodies: TFIIB (clone 2F6A3H4, Cell Signaling-4169, 1:1000), GAPDH (anti-mouse, clone 6C5, Thermofisher-AM4300, 1:5000 or anti-rabbit, clone 14C10, Cell Signaling-2118, 1:1000), p27 (ProteinTech-15086-1-AP, 1:1000), TRIM28 (Cell Signaling-4123, 1:1000), TRIM24 (clone E93TN, Cell Signaling-79030, 1:1000), Vinculin (Abcam-ab91459, 1:1000), FK2 (Enzo-BML-PW8810-0100, 1:1000), NRF2 (ProteinTech-16396-1-AP, 1:000), Caspase-3 (Cell Signaling-9662, 1:1000), Caspase-8 (clone E7, Abcam-ab32397, 1:1000), γH2AX (Bethyl-A300-081A-M, 1:1000), p53 (clone 1C12, Cell Signaling-2524, 1:1000), Goat Anti-Mouse IgG(H+L) Human ads-HRP (Southern Biotech, 1:5,000), Goat Anti-Rabbit IgG(H+L) Human ads-HRP (Southern Biotech, 1:5000), Strep-Tag II Antibody HRP Conjugate (Sigma Aldrich-71591-3, 1:2000).

Techniques: Construct, Western Blot, Marker, Infection, Control, Inhibition, Expressing, Activation Assay, Mutagenesis, Negative Control, Protease Inhibitor

(A) TRExRTA BCBL-1 cells treated with 100 μg/mL Chx and endogenous TFIIB protein were analyzed by western blotting. p27 is a control for translational inhibition and GAPDH probed with mouse antibody and vinculin are loading controls. (B) iSLK-KSHV(+) BAC16 cells were reactivated with doxycycline for the given timepoints prior to western blotting. Vinculin serves as a loading control. Note the lack of appreciable procaspase-3 cleavage and activation. These samples come from the same experiment as . (C) iSLK-KSHV(+) BAC16 cells treated with 250 μg/mL cycloheximide (Chx) for given timepoints. Note lack of procaspase-3 activation.

Journal: bioRxiv

Article Title: The RNA polymerase II general transcription factor TFIIB is a target for transcriptome control during cellular stress and viral infection

doi: 10.1101/2024.01.16.575933

Figure Lengend Snippet: (A) TRExRTA BCBL-1 cells treated with 100 μg/mL Chx and endogenous TFIIB protein were analyzed by western blotting. p27 is a control for translational inhibition and GAPDH probed with mouse antibody and vinculin are loading controls. (B) iSLK-KSHV(+) BAC16 cells were reactivated with doxycycline for the given timepoints prior to western blotting. Vinculin serves as a loading control. Note the lack of appreciable procaspase-3 cleavage and activation. These samples come from the same experiment as . (C) iSLK-KSHV(+) BAC16 cells treated with 250 μg/mL cycloheximide (Chx) for given timepoints. Note lack of procaspase-3 activation.

Techniques: Western Blot, Control, Inhibition, Activation Assay

(A) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStrep protein were pulse-labelled for 18 h with L-HPG, harvested at the indicated timepoints post-chase for strep pulldown, then labelled with picolyl-azide-sulfo-Cy5. For quantification, strep signal was normalized to input signal and the 0 h timepoint and plotted. **** P < 0.0001, * P = 0.0103; one-way ANOVA with Tukey multiple comparison test. (B) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStrep were treated with 100 μg/mL cycloheximide (Chx) and harvested at indicated timepoints for western analysis. p27 is a control for Chx activity and vinculin is a loading control. (C) iSLK-KSHV(+) BAC16 cells were treated with 250 μg/mL Chx, harvested at the given timepoints and western blotted for the indicated proteins. Vinculin and GAPDH probed with mouse antibody serve as loading controls. (D) TRExRTA BCBL-1 cells or (E) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStep were treated with 100 μg/mL Chx for 6h with or without 5 μM carfilzomib proteasome inhibitor (carf) and endogenous TFIIB and strep-tagged protein analyzed by western blotting. p53 and p27 are positive controls for Chx activity and FK2 marks poly-ubiquitylated protein as a positive control for proteasomal inhibition; GAPDH probed with mouse antibody is a loading control. (F) TRExRTA BCBL-1 cells treated with 100 μg/mL Chx for 8 h with or without 10 μM TAK-243, a UBA1-targetting inhibitor of ubiquitylation, and western blotted with the indicated antibodies. NRF2 is a control for translation and ubiquitylation inhibition, and GAPDH probed with rabbit antibody is a loading control.

Journal: bioRxiv

Article Title: The RNA polymerase II general transcription factor TFIIB is a target for transcriptome control during cellular stress and viral infection

doi: 10.1101/2024.01.16.575933

Figure Lengend Snippet: (A) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStrep protein were pulse-labelled for 18 h with L-HPG, harvested at the indicated timepoints post-chase for strep pulldown, then labelled with picolyl-azide-sulfo-Cy5. For quantification, strep signal was normalized to input signal and the 0 h timepoint and plotted. **** P < 0.0001, * P = 0.0103; one-way ANOVA with Tukey multiple comparison test. (B) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStrep were treated with 100 μg/mL cycloheximide (Chx) and harvested at indicated timepoints for western analysis. p27 is a control for Chx activity and vinculin is a loading control. (C) iSLK-KSHV(+) BAC16 cells were treated with 250 μg/mL Chx, harvested at the given timepoints and western blotted for the indicated proteins. Vinculin and GAPDH probed with mouse antibody serve as loading controls. (D) TRExRTA BCBL-1 cells or (E) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStep were treated with 100 μg/mL Chx for 6h with or without 5 μM carfilzomib proteasome inhibitor (carf) and endogenous TFIIB and strep-tagged protein analyzed by western blotting. p53 and p27 are positive controls for Chx activity and FK2 marks poly-ubiquitylated protein as a positive control for proteasomal inhibition; GAPDH probed with mouse antibody is a loading control. (F) TRExRTA BCBL-1 cells treated with 100 μg/mL Chx for 8 h with or without 10 μM TAK-243, a UBA1-targetting inhibitor of ubiquitylation, and western blotted with the indicated antibodies. NRF2 is a control for translation and ubiquitylation inhibition, and GAPDH probed with rabbit antibody is a loading control.

Techniques: Expressing, Comparison, Western Blot, Control, Activity Assay, Positive Control, Inhibition

(A) iSLK-KSHV(+) BAC16 cells +/-reactivation for 36 h, treated with 250 μg/mL cycloheximide (Chx) for the indicated timepoints, then western blotted with the indicated antibodies. KSHV SOX is a control for infection, p27 is a control for translation inhibition, and GAPDH probed with rabbit antibody is a loading control. (B) TRExRTA BCBL-1 cells or (C) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStrep were mock-treated or reactivated for 18 h, treated with 100 μg/mL Chx, and western blotted for the indicated proteins. Vinculin is a loading control. (D) Western blots of TRExRTA BCBL-1 cells treated with 18 h of TRIM28 or nontargeting control (siNTC) siRNAs, followed by 100 μg/mL Chx treatment for the indicated times. (E) Western blots of TRExRTA BCBL-1 cells +/-18 h reactivation and treatment with 1mM of the viral DNA replication inhibitor phosphonoacetic acid (PAA) followed by a timecourse with 100 μg/mL Chx. Early viral gene SOX expression is not affected by inhibition of DNA replication, but late gene K8.1 is dependent on DNA replication and is thus not expressed. Black arrow indicates appropriate p27 band under non-specific antibody signal. GAPDH probed with mouse antibody is a loading control.

Journal: bioRxiv

Article Title: The RNA polymerase II general transcription factor TFIIB is a target for transcriptome control during cellular stress and viral infection

doi: 10.1101/2024.01.16.575933

Figure Lengend Snippet: (A) iSLK-KSHV(+) BAC16 cells +/-reactivation for 36 h, treated with 250 μg/mL cycloheximide (Chx) for the indicated timepoints, then western blotted with the indicated antibodies. KSHV SOX is a control for infection, p27 is a control for translation inhibition, and GAPDH probed with rabbit antibody is a loading control. (B) TRExRTA BCBL-1 cells or (C) TRExRTA BCBL-1 cells expressing TFIIB(D207A)-2xStrep were mock-treated or reactivated for 18 h, treated with 100 μg/mL Chx, and western blotted for the indicated proteins. Vinculin is a loading control. (D) Western blots of TRExRTA BCBL-1 cells treated with 18 h of TRIM28 or nontargeting control (siNTC) siRNAs, followed by 100 μg/mL Chx treatment for the indicated times. (E) Western blots of TRExRTA BCBL-1 cells +/-18 h reactivation and treatment with 1mM of the viral DNA replication inhibitor phosphonoacetic acid (PAA) followed by a timecourse with 100 μg/mL Chx. Early viral gene SOX expression is not affected by inhibition of DNA replication, but late gene K8.1 is dependent on DNA replication and is thus not expressed. Black arrow indicates appropriate p27 band under non-specific antibody signal. GAPDH probed with mouse antibody is a loading control.

Techniques: Western Blot, Control, Infection, Inhibition, Expressing

(A) Western blots from of TRExRTA BCBL-1 cells nucleofected with TRIM24 siRNA or non-targeting control (siNTC) for 18 h followed by a timecourse with 100 μg/mL cycloheximide treatment (Chx). p27 is a translation inhibitor control and GAPDH probed with mouse antibody a loading control. (B) Western blots of iSLK-KSHV(+) BAC16 cells +/-reactivation and treated with 1mM of phosphonoacetic acid (PAA). SOX is a viral early gene and K8.1 is a late gene (expressed only after DNA replication).

Journal: bioRxiv

Article Title: The RNA polymerase II general transcription factor TFIIB is a target for transcriptome control during cellular stress and viral infection

doi: 10.1101/2024.01.16.575933

Figure Lengend Snippet: (A) Western blots from of TRExRTA BCBL-1 cells nucleofected with TRIM24 siRNA or non-targeting control (siNTC) for 18 h followed by a timecourse with 100 μg/mL cycloheximide treatment (Chx). p27 is a translation inhibitor control and GAPDH probed with mouse antibody a loading control. (B) Western blots of iSLK-KSHV(+) BAC16 cells +/-reactivation and treated with 1mM of phosphonoacetic acid (PAA). SOX is a viral early gene and K8.1 is a late gene (expressed only after DNA replication).

Techniques: Western Blot, Control

Review

Structured Review

Images

1) Product Images from "GLS1‐RNA Polymerase II Axis Mediates Glutamine‐Dependent Hepatoprotective Effects on Alcoholic Liver Disease in High‐Protein Diets"

Similar Products

Image Search Results