Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: TNFα selectively activates the IRE1α/XBP1 endoplasmic reticulum stress pathway in human airway smooth muscle cells

doi: 10.1152/ajplung.00212.2019

Figure Lengend Snippet: Exposure to tunicamycin activates the inositol-requiring enzyme1 (IRE1α)/X-box binding protein 1 (XBP1) pathway in airway smooth muscle (ASM) cells. A: 12-h exposure to tunicamycin significantly increased the ratio of phosphorylated to total IRE1α in ASM cells compared with untreated controls. Representative Western blots are shown. B: 12-h exposure to tunicamycin also significantly increased total IRE1α expression [relative to ribosomal protein S16 (RPS16)] in ASM cells compared with untreated controls. Representative Western blots are shown. C: splicing of XBP1 (relative to RPS16) increased significantly after exposure to tunicamycin for 6 and 12 h in ASM cells compared with untreated controls. Representative Western blots are shown. Data are presented as medians and interquartile range (IQR) represented as a box-and-whisker plot. Results were analyzed using a 2-way ANOVA. Untreated controls for each time point were performed. *Significant difference (P < 0.05) compared with untreated control (n = 5 patients).

Article Snippet: Total IRE1α (Cell Signaling Technology Cat no. 3294, RRID:AB_823545) and total PERK (Cell Signaling Technology Cat no. 3192, RRID:AB_2095847) were detected with specific antibodies obtained from Cell Signaling (Danvers, MA).

Techniques: Binding Assay, Western Blot, Expressing, Whisker Assay

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: TNFα selectively activates the IRE1α/XBP1 endoplasmic reticulum stress pathway in human airway smooth muscle cells

doi: 10.1152/ajplung.00212.2019

Figure Lengend Snippet: TNFα activates the inositol-requiring enzyme1 (IRE1α)/X-box binding protein 1 (XBP1) pathway in airway smooth muscle (ASM) cells in a concentration-dependent fashion. A: exposure to TNFα at 20, 50, and 100 ng/mL (12-h) significantly increased the ratio of phosphorylated (p) to total IRE1α in ASM cells compared with untreated controls. Representative Western blots are shown. B: exposure to TNFα did not change total IRE1α expression [relative to ribosomal protein S16 (RPS16)} in ASM cells compared with untreated controls. Representative Western blots are shown. C: splicing of XBP1 (relative to RPS16) increased significantly after exposure to TNFα at 50 and 100 ng/mL in ASM cells compared with untreated controls. Representative Western blots are shown. Data are presented as medians and interquartile range (IQR) represented as a box-and-whisker plot. Results were analyzed using a 2-way ANOVA. *Significant difference (P < 0.05) compared with untreated control (n = 5 patients).

Article Snippet: Total IRE1α (Cell Signaling Technology Cat no. 3294, RRID:AB_823545) and total PERK (Cell Signaling Technology Cat no. 3192, RRID:AB_2095847) were detected with specific antibodies obtained from Cell Signaling (Danvers, MA).

Techniques: Binding Assay, Concentration Assay, Western Blot, Expressing, Whisker Assay

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: TNFα selectively activates the IRE1α/XBP1 endoplasmic reticulum stress pathway in human airway smooth muscle cells

doi: 10.1152/ajplung.00212.2019

Figure Lengend Snippet: TNFα activates the inositol-requiring enzyme1 (IRE1α)/X-box binding protein 1 (XBP1) pathway in airway smooth muscle (ASM) cells in a time-dependent fashion. A: exposure to TNFα for 6, 12, and 24 h (50 ng/mL) significantly increased the ratio of phosphorylated to total IRE1α in ASM cells compared with untreated controls. Representative Western blots are shown. B: exposure to TNFα at different times did not significantly change total IRE1α expression [relative to ribosomal protein S16 (RPS16)} in ASM cells compared with untreated controls. Representative Western blots are shown. C: splicing of XBP1 (relative to RPS16) increased significantly after exposure to TNFα at 12 and 24 h (50 ng/mL) in ASM cells compared with untreated controls. Representative Western blots are shown. Data are presented as medians and interquartile range (IQR) represented as a box-and-whisker plot. Results were analyzed using a 2-way ANOVA. Untreated controls for each time point were performed.

Article Snippet: Total IRE1α (Cell Signaling Technology Cat no. 3294, RRID:AB_823545) and total PERK (Cell Signaling Technology Cat no. 3192, RRID:AB_2095847) were detected with specific antibodies obtained from Cell Signaling (Danvers, MA).

Techniques: Binding Assay, Western Blot, Expressing, Whisker Assay

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: TNFα selectively activates the IRE1α/XBP1 endoplasmic reticulum stress pathway in human airway smooth muscle cells

doi: 10.1152/ajplung.00212.2019

Figure Lengend Snippet: Tempol reduces TNFα-induced increase in inositol-requiring enzyme1 (IRE1α) phosphorylation and reduction in mitofusin 2 (Mfn2) expression in airway smooth muscle (ASM) cells compared with untreated control ASM cells. A: exposure to TNFα for 12 h (50 ng/mL) significantly increased the ratio of phosphorylated to total IRE1α in ASM cells compared with untreated controls, while concomitant exposure with antioxidant tempol reduced the effect of TNFα. Representative Western blots are shown. B: exposure to TNFα for 12 h (50 ng/mL) significantly decreased Mfn2 expression [relative to ribosomal protein S16 (RPS16)] in ASM cells compared with untreated controls, while concomitant exposure with antioxidant tempol reduced the effect of TNFα. Representative Western blots are shown. Data are presented as medians and interquartile range (IQR) represented as a box-and-whisker plot. Results were analyzed using a 2-way ANOVA. *Significant difference (P < 0.05) compared with untreated control (n = 5 patients).

Article Snippet: Total IRE1α (Cell Signaling Technology Cat no. 3294, RRID:AB_823545) and total PERK (Cell Signaling Technology Cat no. 3192, RRID:AB_2095847) were detected with specific antibodies obtained from Cell Signaling (Danvers, MA).

Techniques: Expressing, Western Blot, Whisker Assay

Journal: Life Science Alliance

Article Title: Paneth cell α-defensin misfolding correlates with dysbiosis and ileitis in Crohn’s disease model mice

doi: 10.26508/lsa.201900592

Figure Lengend Snippet: (A, B) Representative transmission electron microscopy images of Paneth cells at the base of ileal crypts in (A) ICR and (B) SAMP1/YitFc mice. Scale bars indicate 2 μm. (C, D) Quantitative analysis of (C) granule number and (D) ER lumen diameter in Paneth cells (n = 3/each week for SAMP1/YitFc mice). For the measurements, three Paneth cells were randomly selected from each mouse. (E) SDS–PAGE Western blot analysis of ER stress markers, pIRE1α, ATF4, cleaved-ATF6, and GRP78 in ileal crypts (n = 4/each group). Total-IRE1α and HPRT1 was used as loading control. (F) Relative expression level of ER stress markers calculated from the band intensity. Error bars represent mean ± SEM. (C, D, F) Statistical significance was evaluated by t test in (C, D), and one-way ANOVA followed by Tukey’s post hoc test in (F). P < 0.05 was considered statistically significant. * P < 0.05, † P < 0.01, § P < 0.001. E, ER; G, granules; N, nucleus; n.s., not significant.

Article Snippet: After transfer, the membranes were blocked by 5% BSA (Sigma-Aldrich) in 0.1% TBS-T at 4°C for overnight and then reacted with the following primary antibodies: anti-ATF4 (1/1,000, #11815; Cell Signaling Technology), anti-phospho-IRE1α (1 μg/ml, NB100-2323; Novus Biologicals), anti-total IRE1α (1 μg/ml, NB100-2324; Novus Biologicals), anti-ATF6 (1 μg/ml, NBP1-40256; Novus Biologicals), anti-GRP78 (1/1,000, #3177; Cell Signaling Technology), and anti-HPRT1 (0.018 μg/ml, ab109021; Abcam) at 4°C overnight.

Techniques: Transmission Assay, Electron Microscopy, SDS Page, Western Blot, Control, Expressing

Journal: Cell reports

Article Title: IRE1α promotes phagosomal calcium flux to enhance macrophage fungicidal activity.

doi: 10.1016/j.celrep.2025.115694

Figure Lengend Snippet: Figure 1. Candida albicans infection results in activation of macrophage IRE1α (A) iBMDM cell lines (WT or IRE1ΔR) were infected with C. albicans (Ca), treated with LPS, thapsi- gargin, or mock for 4 h. Xbp1 mRNA splicing was measured by semi-quantitative RT-PCR amplifi- cation of the Xbp1 transcript, followed by treat- ment with PstI, which recognizes a cleavage site within the 26-bp intron that is removed by IRE1α, resulting in cleavage of the unspliced isoform, specifically. (B) Immunoblot analysis of lysates from WT or IRE1ΔR iBMDM to confirm IRE1α truncation in IRE1ΔR cells resulting from removal of floxed exons 20 and 21. (C) Expression of spliced Xbp1 over a time course following Ca infection. (D) Expression of spliced Xbp1 at 4 h post-infec- tion (hpi) with the indicated isolates. (E) Expression of spliced Xbp1 at 4 h following Ca infection or LPS treatment of WT or CARD9 KO iBMDM. (F) Expression of spliced Xbp1 at 4 h following Ca infection and depleted zymosan (d-zymosan) or LPS treatment of WT or IRE1ΔR iBMDM. (G) Expression of spliced Xbp1 at 4 h following Ca infection, LPS treatment, or d-zymosan treatment of WT or TLR2/4/9 KO iBMDM. (H) Expression of spliced Xbp1 at 4 h following Ca infection or LPS treatment of two pairs of clonal iBMDM (WT or TRAF6 KO). Closed symbols are data from WT-1 and KO-1; open symbols are data from WT-2 and KO-2. Data are representative of 3–4 individual experiments. Graphs show the mean ± SEM of biological repli- cates (C–H). For all experiments, Ca MOI = 1, LPS treatment = 100 ng/mL, d-zymosan = 100 μg/mL, thapsigargin = 5 μM. Statistical analysis details are in STAR Methods.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-IRE1α Cell Signaling Cat#3924; RRID:AB_823545 Rabbit polyclonal anti-XBP1 Abcam Cat#ab37152; RRID:AB_778939 Rabbit polyclonal anti-CARD9 Cell Signaling Cat#12283; RRID:AB_2797869 Mouse monoclonal anti-ACTIN ThermoFisher Cat#MA5-11869; RRID:AB_11004139 Rabbit monoclonal anti-TRAF6 Abcam Cat#ab40675; RRID:AB_778573 Rabbit polyclonal anti-K48-linkage specific polyubiquitin Cell Signaling Cat#4289; RRID:AB_10557239 Rabbit monoclonal anti-ATF6 Cell Signaling Cat#65880; RRID:AB_2799696 Rabbit polyclonal anti-phosphoeIF2α Ser51 Cell Signaling Cat#9721; RRID:AB_330951 Mouse monoclonal anti- eIF2α Cell Signaling Cat#2103; RRID:AB_836874 Rabbit monoclonal anti-ATF4 Cell Signaling Cat#11815; RRID:AB_2616025 Rabbit polyclonal anti-Candida albicans-FITC LSBio Cat#LS-C103355 Rat monoclonal anti-LAMP1 DSHB Cat#1D4B; RRID:AB_2134500 Mouse monoclonal anti-EEA1 ThermoFisher Cat#MA5-31575 Rabbit monoclonal anti-KDEL Abcam Cat#ab176333; RRID:AB_2819147 Rat monoclonal anti-CD11b, BV421-conjugated Biolegend Cat#414-0112-82 Bacterial and virus strains MRSA-mCherry Methicillin-resistant Staphyloccocus aureus USA300 LAC harboring pSarA-mCherry plasmid Boles et al. 200873 N/A Chemicals, peptides, and recombinant proteins Thioflavin T Cayman Chemical Cat#32553; CAS Number 2390-54-7 TRIzol reagent Thermo Fisher Cat#15596018 Turbo DNase I Thermo Fisher Cat#AM2238 Paraformaldehyde Electron Microscopy Services Cat#15700 Calcofluor white Sigma-Aldrich Cat#18909 Bovine serum albumin Thermo Fisher Cat#BP9706-100 Sulforhodamine B Sigma-Aldrich Cat#230162; CAS Number 3520-42-1 Critical commercial assays Lookout Mycoplasma PCR Detection Kit Sigma-Aldrich Cat#MP0035 iScript cDNA synthesis kit BioRad Cat# 1708891 PowerUP SYBR Green qPCR master mix Thermo Fisher Cat# A25742 Fluo-4 Calcium Imaging Kit Invitrogen Cat#F10489 LysoSensor Yellow/Blue DND-160 Thermo Fisher Cat#L7545 LysoTracker Deep Red Invitrogen Cat#L12492 Duolink In Situ PLA Kit Sigma-Aldrich Cat#DUO92008 Cat#DUO92002 Cat#DUO92004 Fluorospheres Thermo Fisher Cat#F8801 (Continued on next page) Cell Reports 44, 115694, May 27, 2025 15

Techniques: Infection, Activation Assay, Quantitative RT-PCR, Western Blot, Expressing

Journal: Cell reports

Article Title: IRE1α promotes phagosomal calcium flux to enhance macrophage fungicidal activity.

doi: 10.1016/j.celrep.2025.115694

Figure Lengend Snippet: Figure 4. IRE1α promotes phagosome maturation during C. albicans infection (A) Representative images showing LAMP1 (yellow) recruitment to phagosomes containing iRFP-expressing C. albicans (magenta) in IRE1 WT or IRE1ΔR iBMDM at indicated times post-infection. (B) Quantification of LAMP1 recruitment to phagosomes containing C. albicans in IRE1 WT or IRE1ΔR iBMDM, as measured by LAMP1 mean fluorescence in- tensity associated with C. albicans-expressed iRFP. (C) Quantification of C. albicans killing in IRE1 WT and IRE1ΔR macrophages treated with BafA or DMSO control.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-IRE1α Cell Signaling Cat#3924; RRID:AB_823545 Rabbit polyclonal anti-XBP1 Abcam Cat#ab37152; RRID:AB_778939 Rabbit polyclonal anti-CARD9 Cell Signaling Cat#12283; RRID:AB_2797869 Mouse monoclonal anti-ACTIN ThermoFisher Cat#MA5-11869; RRID:AB_11004139 Rabbit monoclonal anti-TRAF6 Abcam Cat#ab40675; RRID:AB_778573 Rabbit polyclonal anti-K48-linkage specific polyubiquitin Cell Signaling Cat#4289; RRID:AB_10557239 Rabbit monoclonal anti-ATF6 Cell Signaling Cat#65880; RRID:AB_2799696 Rabbit polyclonal anti-phosphoeIF2α Ser51 Cell Signaling Cat#9721; RRID:AB_330951 Mouse monoclonal anti- eIF2α Cell Signaling Cat#2103; RRID:AB_836874 Rabbit monoclonal anti-ATF4 Cell Signaling Cat#11815; RRID:AB_2616025 Rabbit polyclonal anti-Candida albicans-FITC LSBio Cat#LS-C103355 Rat monoclonal anti-LAMP1 DSHB Cat#1D4B; RRID:AB_2134500 Mouse monoclonal anti-EEA1 ThermoFisher Cat#MA5-31575 Rabbit monoclonal anti-KDEL Abcam Cat#ab176333; RRID:AB_2819147 Rat monoclonal anti-CD11b, BV421-conjugated Biolegend Cat#414-0112-82 Bacterial and virus strains MRSA-mCherry Methicillin-resistant Staphyloccocus aureus USA300 LAC harboring pSarA-mCherry plasmid Boles et al. 200873 N/A Chemicals, peptides, and recombinant proteins Thioflavin T Cayman Chemical Cat#32553; CAS Number 2390-54-7 TRIzol reagent Thermo Fisher Cat#15596018 Turbo DNase I Thermo Fisher Cat#AM2238 Paraformaldehyde Electron Microscopy Services Cat#15700 Calcofluor white Sigma-Aldrich Cat#18909 Bovine serum albumin Thermo Fisher Cat#BP9706-100 Sulforhodamine B Sigma-Aldrich Cat#230162; CAS Number 3520-42-1 Critical commercial assays Lookout Mycoplasma PCR Detection Kit Sigma-Aldrich Cat#MP0035 iScript cDNA synthesis kit BioRad Cat# 1708891 PowerUP SYBR Green qPCR master mix Thermo Fisher Cat# A25742 Fluo-4 Calcium Imaging Kit Invitrogen Cat#F10489 LysoSensor Yellow/Blue DND-160 Thermo Fisher Cat#L7545 LysoTracker Deep Red Invitrogen Cat#L12492 Duolink In Situ PLA Kit Sigma-Aldrich Cat#DUO92008 Cat#DUO92002 Cat#DUO92004 Fluorospheres Thermo Fisher Cat#F8801 (Continued on next page) Cell Reports 44, 115694, May 27, 2025 15

Techniques: Infection, Expressing, Fluorescence, Control

Journal: Cell reports

Article Title: IRE1α promotes phagosomal calcium flux to enhance macrophage fungicidal activity.

doi: 10.1016/j.celrep.2025.115694

Figure Lengend Snippet: Figure 5. IRE1α promotes phagosomal calcium flux and contact sites between the ER and endosomal network (A) Representative micrographs of WT or IRE1ΔR iBMDM following phagocytosis of C. albicans (20 min post-infection; MOI 2) showing early cellular calcium flux, and influx of calcium specifically in the phagosome following phagocytosis of C. albicans. Scale bar, 10 μm. (B) Quantification of calcium-high phagosomes, defined by a 1.25-fold increase of the mean fluorescence intensity of the cell (20 min post-infection; MOI 2). (C) Violin plot of the ratio of phagosomal to cytosolic mean fluorescence intensity of Fluo4 (20 min post-infection; MOI 2). (D) Quantification of LAMP1 recruitment to phagosomes containing C. albicans in IRE1 WT or IRE1ΔR iBMDM with or without treatment of BAPTA-AM (calcium chelator), as measured by LAMP1 mean fluorescence intensity associated with C. albicans-expressed iRFP. (E) Representative confocal micrographs of PLA puncta representing sites of proximity between the ER and early endosomes/phagosomes at resting state (mock) or during C. albicans infection. Scale bar, 10 μm. (F) Quantification of PLA puncta in IRE1 WT or IRE1ΔR iBMDM at resting state (mock). Values are the mean ± SEM from 3 to 4 biological replicates, as indicated by data points. Statistical analysis details are in STAR Methods.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-IRE1α Cell Signaling Cat#3924; RRID:AB_823545 Rabbit polyclonal anti-XBP1 Abcam Cat#ab37152; RRID:AB_778939 Rabbit polyclonal anti-CARD9 Cell Signaling Cat#12283; RRID:AB_2797869 Mouse monoclonal anti-ACTIN ThermoFisher Cat#MA5-11869; RRID:AB_11004139 Rabbit monoclonal anti-TRAF6 Abcam Cat#ab40675; RRID:AB_778573 Rabbit polyclonal anti-K48-linkage specific polyubiquitin Cell Signaling Cat#4289; RRID:AB_10557239 Rabbit monoclonal anti-ATF6 Cell Signaling Cat#65880; RRID:AB_2799696 Rabbit polyclonal anti-phosphoeIF2α Ser51 Cell Signaling Cat#9721; RRID:AB_330951 Mouse monoclonal anti- eIF2α Cell Signaling Cat#2103; RRID:AB_836874 Rabbit monoclonal anti-ATF4 Cell Signaling Cat#11815; RRID:AB_2616025 Rabbit polyclonal anti-Candida albicans-FITC LSBio Cat#LS-C103355 Rat monoclonal anti-LAMP1 DSHB Cat#1D4B; RRID:AB_2134500 Mouse monoclonal anti-EEA1 ThermoFisher Cat#MA5-31575 Rabbit monoclonal anti-KDEL Abcam Cat#ab176333; RRID:AB_2819147 Rat monoclonal anti-CD11b, BV421-conjugated Biolegend Cat#414-0112-82 Bacterial and virus strains MRSA-mCherry Methicillin-resistant Staphyloccocus aureus USA300 LAC harboring pSarA-mCherry plasmid Boles et al. 200873 N/A Chemicals, peptides, and recombinant proteins Thioflavin T Cayman Chemical Cat#32553; CAS Number 2390-54-7 TRIzol reagent Thermo Fisher Cat#15596018 Turbo DNase I Thermo Fisher Cat#AM2238 Paraformaldehyde Electron Microscopy Services Cat#15700 Calcofluor white Sigma-Aldrich Cat#18909 Bovine serum albumin Thermo Fisher Cat#BP9706-100 Sulforhodamine B Sigma-Aldrich Cat#230162; CAS Number 3520-42-1 Critical commercial assays Lookout Mycoplasma PCR Detection Kit Sigma-Aldrich Cat#MP0035 iScript cDNA synthesis kit BioRad Cat# 1708891 PowerUP SYBR Green qPCR master mix Thermo Fisher Cat# A25742 Fluo-4 Calcium Imaging Kit Invitrogen Cat#F10489 LysoSensor Yellow/Blue DND-160 Thermo Fisher Cat#L7545 LysoTracker Deep Red Invitrogen Cat#L12492 Duolink In Situ PLA Kit Sigma-Aldrich Cat#DUO92008 Cat#DUO92002 Cat#DUO92004 Fluorospheres Thermo Fisher Cat#F8801 (Continued on next page) Cell Reports 44, 115694, May 27, 2025 15

Techniques: Infection, Fluorescence

Journal: PLoS ONE

Article Title: HCV Causes Chronic Endoplasmic Reticulum Stress Leading to Adaptation and Interference with the Unfolded Protein Response

doi: 10.1371/journal.pone.0024660

Figure Lengend Snippet: HuH7.5.1 hepatoma cells were infected with HCV-JFH1. Following infection, protein was extracted at the indicated time points. The expression of (a) phospho-IRE1, and (b) cleaved and non-cleavedATF6 (c) phospho-eIF2α were analyzed by western blotting. (d) CHOP translocation to the nucleus was assessed by immunofluorecence in control HuH7.5.1 cells or JFH1 infected cells on day 5, isotype anti-rabbit IgG was used as control. Results are means±S.D. of 3 experiments. Blots and picture are from a representative experiment (see also ).

Article Snippet: The following antibodies were used: NS5a (Virogen, MA, USA #276-A), XBP-1 (Santa Cruz CA, #7160), total-IRE1 α (Cell Signaling #3294) phospho-IRE1α (Abcam, Cambridge, UK, #48187), phospho/total- eiF2α (Cell Signaling #9721, #9722), BiP (Abcam #21685), ATF6 (Abcam #11909), CHOP (Santa-Cruz, #sc-575).

Techniques: Infection, Expressing, Western Blot, Translocation Assay, Control

Journal: PLoS ONE

Article Title: HCV Causes Chronic Endoplasmic Reticulum Stress Leading to Adaptation and Interference with the Unfolded Protein Response

doi: 10.1371/journal.pone.0024660

Figure Lengend Snippet: HCV infected cells were stimulated by decreasing doses of thapsigargin at day 5 and at day 14 post infection and the effect of stimulation on (a) XBP-1 splicing or (b) IRE1 and (c) eIF2α phosphorylation was assessed by RT-PCR and immunoblotting respectively. Quantification of the results was obtained from densitometric scanning of the blots (means±S.D.) of 3 experiments. * = p<0.05 (ANOVA, one way) for day 5 versus day 14 post infection.

Article Snippet: The following antibodies were used: NS5a (Virogen, MA, USA #276-A), XBP-1 (Santa Cruz CA, #7160), total-IRE1 α (Cell Signaling #3294) phospho-IRE1α (Abcam, Cambridge, UK, #48187), phospho/total- eiF2α (Cell Signaling #9721, #9722), BiP (Abcam #21685), ATF6 (Abcam #11909), CHOP (Santa-Cruz, #sc-575).

Techniques: Infection, Phospho-proteomics, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Cell Death & Disease

Article Title: Characterization of the role of TMEM45A in cancer cell sensitivity to cisplatin

doi: 10.1038/s41419-019-2088-x

Figure Lengend Snippet: Antibodies used for immunofluorescence labeling and western blot.

Article Snippet: IRE1α total , Anti-IRE1α (Rabbit, Novus bilogicals, NB100-2324, 1/1000) , IRDye 800CW (Anti-rabbit, Li-Cor Biosciences 926–32211, 1/10,000).

Techniques: Immunofluorescence, Labeling, Western Blot

Journal: PLoS ONE

Article Title: HCV Causes Chronic Endoplasmic Reticulum Stress Leading to Adaptation and Interference with the Unfolded Protein Response

doi: 10.1371/journal.pone.0024660

Figure Lengend Snippet: HuH7.5.1 hepatoma cells were infected with HCV-JFH1. Following infection, protein was extracted at the indicated time points. The expression of (a) phospho-IRE1, and (b) cleaved and non-cleavedATF6 (c) phospho-eIF2α were analyzed by western blotting. (d) CHOP translocation to the nucleus was assessed by immunofluorecence in control HuH7.5.1 cells or JFH1 infected cells on day 5, isotype anti-rabbit IgG was used as control. Results are means±S.D. of 3 experiments. Blots and picture are from a representative experiment (see also ).

Article Snippet: The following antibodies were used: NS5a (Virogen, MA, USA #276-A), XBP-1 (Santa Cruz CA, #7160), total-IRE1 α (Cell Signaling #3294) phospho-IRE1α (Abcam, Cambridge, UK, #48187), phospho/total- eiF2α (Cell Signaling #9721, #9722), BiP (Abcam #21685), ATF6 (Abcam #11909), CHOP (Santa-Cruz, #sc-575).

Techniques: Infection, Expressing, Western Blot, Translocation Assay, Control

Journal: PLoS ONE

Article Title: HCV Causes Chronic Endoplasmic Reticulum Stress Leading to Adaptation and Interference with the Unfolded Protein Response

doi: 10.1371/journal.pone.0024660

Figure Lengend Snippet: HCV infected cells were stimulated by decreasing doses of thapsigargin at day 5 and at day 14 post infection and the effect of stimulation on (a) XBP-1 splicing or (b) IRE1 and (c) eIF2α phosphorylation was assessed by RT-PCR and immunoblotting respectively. Quantification of the results was obtained from densitometric scanning of the blots (means±S.D.) of 3 experiments. * = p<0.05 (ANOVA, one way) for day 5 versus day 14 post infection.

Article Snippet: The following antibodies were used: NS5a (Virogen, MA, USA #276-A), XBP-1 (Santa Cruz CA, #7160), total-IRE1 α (Cell Signaling #3294) phospho-IRE1α (Abcam, Cambridge, UK, #48187), phospho/total- eiF2α (Cell Signaling #9721, #9722), BiP (Abcam #21685), ATF6 (Abcam #11909), CHOP (Santa-Cruz, #sc-575).

Techniques: Infection, Phospho-proteomics, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Molecular cancer therapeutics

Article Title: Suppression of Nuclear Factor-κB by Glucocorticoid Receptor Blocks Estrogen-induced Apoptosis in Estrogen-deprived Breast Cancer Cells

doi: 10.1158/1535-7163.MCT-18-1363

Figure Lengend Snippet: A, Regulation signaling pathways detected by RPPA. MCF-7:5C cells were treated with E2 (1 nM), Dex (0.1 μM), or a combination of them for different times as indicated. Next, cell lysates were harvested for RPPA analysis. B-E, Validation of signaling pathways regulated by E2 and Dex. MCF-7:5C cells were treated with E2 (1 nM), Dex (0.1 μM), or a combination of them for 24 and 72 hours. Cell lysates were then harvested. Expression of p-Akt and total Akt (B), p-PDK1 and total PDK1 (C), p-p38 and total p38 (D), p-STAT3 and total STAT3 (E) was measured by Western blotting.

Article Snippet: For Western blotting, antibodies against total eIF2α, phosphorylated eIF2α, IRE-1α, total Akt, phosphorylated Akt, total PDK1, phosphorylated PDK1, total STAT3, phosphorylated STAT3, NF-κB p65, NF-κB p105, cleaved PARP, and caspase-7 were obtained from Cell Signaling Technology.

Techniques: Expressing, Western Blot