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Addgene inc encoding scrambled shctrl

Figure 2. H2O2-induced apoptosis of ovarian cancer cells after downregulation <t>of</t> <t>PHLDA1.</t> (A and B) Flow cytometric analysis of 2008 cells (A) and SKOV3 cells (B) expressing control <t>(shctrl)</t> or PHLDA1-targeting shRNAs (shPHLDA1). Lower right and upper right quadrants showed early apoptotic and late apoptotic/necrotic cells, respectively. n=3, *P<0.05, **P<0.001, compared with the shctrl group. (C and D) Western blot analysis of the apoptosis marker c-PARP1 in 2008 (C) and SKOV3 (D) cell lines expressing shctrl or shPHLDA1.

Encoding Scrambled Shctrl, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/plasmids+encoding+shctrl/pLKO%2E1+shControl+(Plasmid+%2374972)/pm34405011-64-7-10
Average 93 stars, based on 19 article reviews

encoding scrambled shctrl - by Bioz Stars, 2026-11

93/100 stars

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1) Product Images from "PHLDA1 Modulates the Endoplasmic Reticulum Stress Response and is required for Resistance to Oxidative Stress-induced Cell Death in Human Ovarian Cancer Cells."

Article Title: PHLDA1 Modulates the Endoplasmic Reticulum Stress Response and is required for Resistance to Oxidative Stress-induced Cell Death in Human Ovarian Cancer Cells.

Journal: Journal of Cancer

doi: 10.7150/jca.45262

Figure 2. H2O2-induced apoptosis of ovarian cancer cells after downregulation of PHLDA1. (A and B) Flow cytometric analysis of 2008 cells (A) and SKOV3 cells (B) expressing control (shctrl) or PHLDA1-targeting shRNAs (shPHLDA1). Lower right and upper right quadrants showed early apoptotic and late apoptotic/necrotic cells, respectively. n=3, *P<0.05, **P<0.001, compared with the shctrl group. (C and D) Western blot analysis of the apoptosis marker c-PARP1 in 2008 (C) and SKOV3 (D) cell lines expressing shctrl or shPHLDA1.

Figure Legend Snippet: Figure 2. H2O2-induced apoptosis of ovarian cancer cells after downregulation of PHLDA1. (A and B) Flow cytometric analysis of 2008 cells (A) and SKOV3 cells (B) expressing control (shctrl) or PHLDA1-targeting shRNAs (shPHLDA1). Lower right and upper right quadrants showed early apoptotic and late apoptotic/necrotic cells, respectively. n=3, *P<0.05, **P<0.001, compared with the shctrl group. (C and D) Western blot analysis of the apoptosis marker c-PARP1 in 2008 (C) and SKOV3 (D) cell lines expressing shctrl or shPHLDA1.

Techniques Used: Expressing, Control, Western Blot, Marker

Figure 3. ER stress-induced apoptosis of ovarian cancer cells after downregulation of PHLDA1. Apoptosis assay of 2008 (A) and SKOV3 (B) cells expressing shctrl or shPHLDA1 after treatment with thapsigargin (Tg). **P<0.001 compared with the shctrl group. Mean ± SD, n = 3.

Figure Legend Snippet: Figure 3. ER stress-induced apoptosis of ovarian cancer cells after downregulation of PHLDA1. Apoptosis assay of 2008 (A) and SKOV3 (B) cells expressing shctrl or shPHLDA1 after treatment with thapsigargin (Tg). **P<0.001 compared with the shctrl group. Mean ± SD, n = 3.

Techniques Used: Apoptosis Assay, Expressing

Figure 5. Expression of autophagy-related proteins, anti-apoptosis proteins, ER stress-associated proteins after PHLDA1-downregulation in ovarian cancer cells. (A) Western blot analysis of Autophagy-related proteins (Beclin-1, P62, and LC3) and anti-apoptosis protein (Bcl-2) in 2008 cells expressing shctrl or shPHLDA1 after treatment with 0.2 mM H2O2. (B and C) Western blot analysis of ER stress-associated proteins (IRE1α, PERK, BIP, ERO1-Lα, and PDI) in 2008 (B) and SKOV3 (C) cells after treatment with 0.2 mM H2O2.

Figure Legend Snippet: Figure 5. Expression of autophagy-related proteins, anti-apoptosis proteins, ER stress-associated proteins after PHLDA1-downregulation in ovarian cancer cells. (A) Western blot analysis of Autophagy-related proteins (Beclin-1, P62, and LC3) and anti-apoptosis protein (Bcl-2) in 2008 cells expressing shctrl or shPHLDA1 after treatment with 0.2 mM H2O2. (B and C) Western blot analysis of ER stress-associated proteins (IRE1α, PERK, BIP, ERO1-Lα, and PDI) in 2008 (B) and SKOV3 (C) cells after treatment with 0.2 mM H2O2.

Techniques Used: Expressing, Western Blot

Transduction:

Article Title: OPA1 protects intervertebral disc and knee joint health in aged mice by maintaining the structure and metabolic functions of mitochondria
Article Snippet: Primary NP cells from Sprague Dawley (Charles River), were obtained and cultured in antibiotic supplemented Dulbecco’s modified Eagle medium (DMEM) and 10% FBS ( ). .. To explore role of OPA1, lentiviral particle production and viral transduction LV-Sh Opa1 clone #1 (TRCN0000091111 ) and LV-SH Opa1 clone #2 (TRCN0000348537) (Sigma, St. Louis, MO, USA) and pLKO.1 ShCtrl and psPAX2 (#12260) and pMD2G (#12259) (Addgene, Cambridge, MA, USA) were used. ..

Plasmid Preparation:

Article Title: Knockdown of heat shock protein family D member 1 (HSPD1) in lung cancer cell altered secretome profile and cancer-associated fibroblast induction.
Article Snippet: The crosstalk between lung cancer cells and cancer-associated fibroblast (CAF) is pivotal in cancer progression.. Heat shock protein family D member 1 (HSPD1) is a potential prognostic biomarker associated with the tumor microenvironment in lung adenocarcinoma (LUAD).. However, the role of HSPD1 in CAF activation remains unclear.

Article Title: CXCR4/CXCL12 axis promotes lymphatic metastasis in tongue squamous cell carcinoma via PI3K/AKT signaling pathway
Article Snippet: .. Lentiviral vectors expressing CXCR4 shRNA (shCXCR4) or non-targeting control shRNA (shControl) were constructed by cloning the target sequences into the pLKO.1 vector (Addgene, Cat10878). ..

Transfection:

Article Title: Chemical reprogramming of fibroblasts into retinal pigment epithelium cells for vision restoration.
Article Snippet: .. The next day, when the cells reached 70–80% confluence, the 943 constructed vectors (pLKO.1-shAscl1-1, pLKO.1-shAscl1-2, pLKO.1-shOlig2-1, pLKO.1-944 shOlig2-2 and pLKO.1-shControl) were co-transfected with the lentiviral packaging vectors 945 pMD2.G (Addgene, 12259) and psPAX2 (Addgene, 12260) into 293T cells using 946 Lipofectamine 3000 (Invitrogen, L3000015) transfection reagent, following the manufacturer's 947 instructions. .. Viral supernatants were collected at 48- and 72-hours post-transfection, filtered 948 through a 0.45 μm syringe filter to remove cellular debris, and concentrated overnight using 949 the Universal Virus Concentration Kit (Beyotime, C2901S).

Construct:

Control:

shRNA:

Negative Control:

Expressing:

Cloning:

other:

Article Title: A commonly inherited human PCSK9 germline variant drives breast cancer metastasis via LRP1 receptor.
Article Snippet: Article A commonly inherited human PCSK9 germline variant drives breast cancer metastasis via LRP1 receptor

Image Search Results

( A ) In vitro deneddylation assay. CSN deneddylation activity in the presence or absence of 500 nM HSC70. Neddylated or non-neddylated CUL1 were detected by immunoblotting with anti-CUL1 antibody. The differences in the deneddylation activity of CSN were illustrated by estimating the percentage of non-neddylated CUL1 to total CUL1 at each time point based on protein band intensities. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 20 min ( P = 0.014) and 40 min ( P = 0.044). ( B ) In vitro deneddylation assay. CSN deneddylation activity in the presence of 500 nM HSC70 with or without 5 µM NR peptide. The bottom graphs show the mean ± s.e.m. ** P < 0.01 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 20 min ( P = 0.005) and 40 min ( P = 0.0023). ( C ) In vitro pulldown assay. A mixture containing Strep-tagged CSN and GST-tagged HSC70 WT or E175S mutant was subjected to immunoprecipitation with Streptactin Sepharose. ( D ) In vitro deneddylation assay. CSN deneddylation activity in the presence or absence of 500 nM HSC70 E175S. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 40 min ( P = 0.036). ( E ) Immunoblotting analysis of the established HSC70 knockdown (shHSC70) cell line. GAPDH protein was used as an internal control, and bar graphs show the relative expression level of HSC70 in shHSC70 cells to those in shCtrl cells ( n = 6 independent expermeriments), and the ratio of neddylated CUL1 to total CUL1 in shCtrl and shHSC70 cells ( n = 9 independent experiments). The box represents the interquartile range (IQR), with the bottom and top edges indicating the 25th and 75th percentiles, respectively. The horizontal line within the box denotes the median (50th percentile). The whiskers extend to the minimum and maximum values from the box. Data points are shown individually. ( F ) Immunoblotting analysis of shCtrl and shHSC70 cells with or without transient FLAG-HSC70 expression. .

Journal: EMBO Reports

Article Title: HSC70 coordinates COP9 signalosome and SCF ubiquitin ligase activity to enable a prompt stress response

doi: 10.1038/s44319-025-00376-x

Figure Lengend Snippet: ( A ) In vitro deneddylation assay. CSN deneddylation activity in the presence or absence of 500 nM HSC70. Neddylated or non-neddylated CUL1 were detected by immunoblotting with anti-CUL1 antibody. The differences in the deneddylation activity of CSN were illustrated by estimating the percentage of non-neddylated CUL1 to total CUL1 at each time point based on protein band intensities. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 20 min ( P = 0.014) and 40 min ( P = 0.044). ( B ) In vitro deneddylation assay. CSN deneddylation activity in the presence of 500 nM HSC70 with or without 5 µM NR peptide. The bottom graphs show the mean ± s.e.m. ** P < 0.01 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 20 min ( P = 0.005) and 40 min ( P = 0.0023). ( C ) In vitro pulldown assay. A mixture containing Strep-tagged CSN and GST-tagged HSC70 WT or E175S mutant was subjected to immunoprecipitation with Streptactin Sepharose. ( D ) In vitro deneddylation assay. CSN deneddylation activity in the presence or absence of 500 nM HSC70 E175S. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 40 min ( P = 0.036). ( E ) Immunoblotting analysis of the established HSC70 knockdown (shHSC70) cell line. GAPDH protein was used as an internal control, and bar graphs show the relative expression level of HSC70 in shHSC70 cells to those in shCtrl cells ( n = 6 independent expermeriments), and the ratio of neddylated CUL1 to total CUL1 in shCtrl and shHSC70 cells ( n = 9 independent experiments). The box represents the interquartile range (IQR), with the bottom and top edges indicating the 25th and 75th percentiles, respectively. The horizontal line within the box denotes the median (50th percentile). The whiskers extend to the minimum and maximum values from the box. Data points are shown individually. ( F ) Immunoblotting analysis of shCtrl and shHSC70 cells with or without transient FLAG-HSC70 expression. .

Article Snippet: Lentiviral particle production involved transfecting plasmids encoding shCtrl or shHSC70 in the pLKO.1 puromycin resistant vector or pCDH-CMV-PA-HA-CSN3-EF1 purovector into HEK293T cells together with pRSV-Rev (Addgene no. 12253), pMD 2.G (Addgene no. 12259), and pMDL g/p RRE plasmids (Addgene no. 12251) using Lipofectamine 2000.

Techniques: In Vitro, Activity Assay, Western Blot, Mutagenesis, Immunoprecipitation, Knockdown, Control, Expressing

( A ) CHX chase assay of endogenous MCL1 with pharmacological inhibition with YM-01 or VER155008. Each protein level was densitometrically quantified (normalized to 0 min) and shown in the graph below. The bottom graph shows the mean ± s.e.m. * P < 0.05, *** P < 0.001 ( n = 3 independent experiments, respectively; Tukey’s HSD test.). Compared with DMSO treatment, a significant difference was observed at 60 min ( P = 0.014), 120 min ( P = 0.0004), and 180 min ( P = 0.0002) in VER155008 treatment group, and was observed at 60 min ( P = 0.033), 120 min ( P < 0.000087), and 180 min ( P = 0.0001) in YM-01 treatment group. ( B ) CHX chase assays of endogenous MCL-1 with or without knockdown of HSC70. The bottom graph shows the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, shCtrl; n = 4 independent experiments, shHSC70; Welch’s t test). A significant difference was observed at 120 min ( P = 0.039) and 180 min ( P = 0.010). ( C ) CHX chase assays of endogenous MCL-1 in shHSC70 cells with or without transient FLAG-HSC70 expression. The CHX chase assay was started 48 h after transfection. The bottom graph shows the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 60 min ( P = 0.026) and 120 min ( P = 0.046). ( D ) and ( E ) In vivo ubiquitination of endogenous MCL-1 ( D ) and FLAG-MCL-1 ( E ). Unmodified and polyubiquitinated FLAG-MCL-1 or endogenous MCL-1 were detected by immunoblotting with FLAG or MCL-1 antibody (*non-specific band). .

Journal: EMBO Reports

Article Title: HSC70 coordinates COP9 signalosome and SCF ubiquitin ligase activity to enable a prompt stress response

doi: 10.1038/s44319-025-00376-x

Figure Lengend Snippet: ( A ) CHX chase assay of endogenous MCL1 with pharmacological inhibition with YM-01 or VER155008. Each protein level was densitometrically quantified (normalized to 0 min) and shown in the graph below. The bottom graph shows the mean ± s.e.m. * P < 0.05, *** P < 0.001 ( n = 3 independent experiments, respectively; Tukey’s HSD test.). Compared with DMSO treatment, a significant difference was observed at 60 min ( P = 0.014), 120 min ( P = 0.0004), and 180 min ( P = 0.0002) in VER155008 treatment group, and was observed at 60 min ( P = 0.033), 120 min ( P < 0.000087), and 180 min ( P = 0.0001) in YM-01 treatment group. ( B ) CHX chase assays of endogenous MCL-1 with or without knockdown of HSC70. The bottom graph shows the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, shCtrl; n = 4 independent experiments, shHSC70; Welch’s t test). A significant difference was observed at 120 min ( P = 0.039) and 180 min ( P = 0.010). ( C ) CHX chase assays of endogenous MCL-1 in shHSC70 cells with or without transient FLAG-HSC70 expression. The CHX chase assay was started 48 h after transfection. The bottom graph shows the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 60 min ( P = 0.026) and 120 min ( P = 0.046). ( D ) and ( E ) In vivo ubiquitination of endogenous MCL-1 ( D ) and FLAG-MCL-1 ( E ). Unmodified and polyubiquitinated FLAG-MCL-1 or endogenous MCL-1 were detected by immunoblotting with FLAG or MCL-1 antibody (*non-specific band). .

Techniques: Inhibition, Knockdown, Expressing, Transfection, In Vivo, Ubiquitin Proteomics, Western Blot

( A ) CHX chase assay of endogenous CDC25A with or without HSC70 knockdown, or pharmacological inhibition with YM-01. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments; Welch’s t test). Compared with shCtrl cells or DMSO treatment, a significant difference was observed at 15 min ( P = 0.025) and 30 min ( P = 0.045) in shHSC70 cells and at 30 min ( P = 0.042) and 60 min ( P = 0.011) in YM-01 treated cells. ( B ) CHX chase assay of endogenous p27 or c-MYC with or without HSC70 knockdown. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). For p27 protein, a significant difference was observed at 120 min ( P = 0.032). For c-MYC protein, significant difference was observed at 60 min ( P = 0.033) and 180 min ( P = 0.045). ( A , B ) Each protein level was densitometrically quantified (normalized to 0 min) and shown in the graph below.

Journal: EMBO Reports

Article Title: HSC70 coordinates COP9 signalosome and SCF ubiquitin ligase activity to enable a prompt stress response

doi: 10.1038/s44319-025-00376-x

Figure Lengend Snippet: ( A ) CHX chase assay of endogenous CDC25A with or without HSC70 knockdown, or pharmacological inhibition with YM-01. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments; Welch’s t test). Compared with shCtrl cells or DMSO treatment, a significant difference was observed at 15 min ( P = 0.025) and 30 min ( P = 0.045) in shHSC70 cells and at 30 min ( P = 0.042) and 60 min ( P = 0.011) in YM-01 treated cells. ( B ) CHX chase assay of endogenous p27 or c-MYC with or without HSC70 knockdown. The bottom graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). For p27 protein, a significant difference was observed at 120 min ( P = 0.032). For c-MYC protein, significant difference was observed at 60 min ( P = 0.033) and 180 min ( P = 0.045). ( A , B ) Each protein level was densitometrically quantified (normalized to 0 min) and shown in the graph below.

Techniques: Knockdown, Inhibition

( A ) Co-IP in HEK293T cells with or without UV irradiation (800 J/m 2 ) using HSC70 antibody, with IgG serving as a negative control. HEK293T cells were treated with 1 µM MLN4924 for 30 min immediately after UV irradiation where indicated. ( B ) Ultraviolet induced degradation of endogenous CDC25A in shCtrl and shHSC70 cells. Each protein level was densitometrically quantified (normalized to 0 min) and shown in the graph. The graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 15 min ( P = 0.031) and 60 min ( P = 0.041). ( C ) Flow cytometry assessment of the cell cycle. shCtrl or shHSC70 cells were synchronized by double thymidine block and were irradiated with low dose UV irradiation (20 J/m 2 ) immediately after release. Cells were harvested and analyzed by fluorescence-assisted cell sorting (FACS) at the indicated time points after UV irradiation. ( D ) Relative proliferation rate of control or HSC70 knockdown HEK293T cells with or without UV irradiation. Relative cell counts were compared to day 0 in all graphs. The graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 3 days ( P = 0.017) after UV irradiation. ( E ) Protein expression level of endogenous cleaved-PARP after UV irradiation. shCtrl or shHSC70 cells were harvested for western blotting at the indicated time point after UV irradiation. The graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 6 h ( P = 0.027) and 8 h ( P = 0.048) after UV irradiation. ( F ) Caspase3/7 assay. Relative caspase 3/7 activity to those at 0 h were shown in the graph. The graphs show the mean ± s.e.m. *** P < 0.001 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 6 h ( P = 0.0005) after UV irradiation. ( G ) Proposed interplay among HSC70, CSN, and NEDD8-SCF. Under basal conditions, HSC70 mainly interacts with CSN and enhances its deneddylation activity. Under SCF-activated conditions, HSC70 alternatively interacts with substrate-bound NEDD8-SCF, thereby promoting SCF ubiquitination activity. .

Journal: EMBO Reports

Article Title: HSC70 coordinates COP9 signalosome and SCF ubiquitin ligase activity to enable a prompt stress response

doi: 10.1038/s44319-025-00376-x

Figure Lengend Snippet: ( A ) Co-IP in HEK293T cells with or without UV irradiation (800 J/m 2 ) using HSC70 antibody, with IgG serving as a negative control. HEK293T cells were treated with 1 µM MLN4924 for 30 min immediately after UV irradiation where indicated. ( B ) Ultraviolet induced degradation of endogenous CDC25A in shCtrl and shHSC70 cells. Each protein level was densitometrically quantified (normalized to 0 min) and shown in the graph. The graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 15 min ( P = 0.031) and 60 min ( P = 0.041). ( C ) Flow cytometry assessment of the cell cycle. shCtrl or shHSC70 cells were synchronized by double thymidine block and were irradiated with low dose UV irradiation (20 J/m 2 ) immediately after release. Cells were harvested and analyzed by fluorescence-assisted cell sorting (FACS) at the indicated time points after UV irradiation. ( D ) Relative proliferation rate of control or HSC70 knockdown HEK293T cells with or without UV irradiation. Relative cell counts were compared to day 0 in all graphs. The graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 3 days ( P = 0.017) after UV irradiation. ( E ) Protein expression level of endogenous cleaved-PARP after UV irradiation. shCtrl or shHSC70 cells were harvested for western blotting at the indicated time point after UV irradiation. The graphs show the mean ± s.e.m. * P < 0.05 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 6 h ( P = 0.027) and 8 h ( P = 0.048) after UV irradiation. ( F ) Caspase3/7 assay. Relative caspase 3/7 activity to those at 0 h were shown in the graph. The graphs show the mean ± s.e.m. *** P < 0.001 ( n = 3 independent experiments, respectively; Welch’s t test). A significant difference was observed at 6 h ( P = 0.0005) after UV irradiation. ( G ) Proposed interplay among HSC70, CSN, and NEDD8-SCF. Under basal conditions, HSC70 mainly interacts with CSN and enhances its deneddylation activity. Under SCF-activated conditions, HSC70 alternatively interacts with substrate-bound NEDD8-SCF, thereby promoting SCF ubiquitination activity. .

Techniques: Co-Immunoprecipitation Assay, Irradiation, Negative Control, Flow Cytometry, Blocking Assay, Fluorescence, FACS, Control, Knockdown, Expressing, Western Blot, Activity Assay, Ubiquitin Proteomics

( A ) Immunostaining of control or HSC70 knockdown HEK293T cells with or without low-dose UV irradiation (20 J/m 2 ) using γH2AX antibody and Hoechst 33342. Scale bars, 10 µm. ( B ) Quantification of cells with γH2AX positive foci with or without UV irradiation ( n = 6 independent experiments for shCtrl cells with or without UV, respectively; n = 7 or 5 independent experiments for shHSC70 cells with or without UV, respectively; Welch’s t test). The graphs show the mean ± s.e.m. NS not significant.

Journal: EMBO Reports

Article Title: HSC70 coordinates COP9 signalosome and SCF ubiquitin ligase activity to enable a prompt stress response

doi: 10.1038/s44319-025-00376-x

Figure Lengend Snippet: ( A ) Immunostaining of control or HSC70 knockdown HEK293T cells with or without low-dose UV irradiation (20 J/m 2 ) using γH2AX antibody and Hoechst 33342. Scale bars, 10 µm. ( B ) Quantification of cells with γH2AX positive foci with or without UV irradiation ( n = 6 independent experiments for shCtrl cells with or without UV, respectively; n = 7 or 5 independent experiments for shHSC70 cells with or without UV, respectively; Welch’s t test). The graphs show the mean ± s.e.m. NS not significant.

Techniques: Immunostaining, Control, Knockdown, Irradiation

Journal: Journal of Cancer

Article Title: PHLDA1 Modulates the Endoplasmic Reticulum Stress Response and is required for Resistance to Oxidative Stress-induced Cell Death in Human Ovarian Cancer Cells.

doi: 10.7150/jca.45262

Figure Lengend Snippet: Figure 2. H2O2-induced apoptosis of ovarian cancer cells after downregulation of PHLDA1. (A and B) Flow cytometric analysis of 2008 cells (A) and SKOV3 cells (B) expressing control (shctrl) or PHLDA1-targeting shRNAs (shPHLDA1). Lower right and upper right quadrants showed early apoptotic and late apoptotic/necrotic cells, respectively. n=3, *P<0.05, **P<0.001, compared with the shctrl group. (C and D) Western blot analysis of the apoptosis marker c-PARP1 in 2008 (C) and SKOV3 (D) cell lines expressing shctrl or shPHLDA1.

Article Snippet: Cells were infected by lentiviral vectors pLKO.1 encoding scrambled shctrl (Addgene, Cambridge, MA, USA) or one of the three PHLDA1-targeting shRNAs (all from Sigma, St. Louis, MO, USA): shPHLDA1-1 (CCG GCC TAA TCC GTA GTA ATT

Techniques: Expressing, Control, Western Blot, Marker

Journal: Journal of Cancer

Article Title: PHLDA1 Modulates the Endoplasmic Reticulum Stress Response and is required for Resistance to Oxidative Stress-induced Cell Death in Human Ovarian Cancer Cells.

doi: 10.7150/jca.45262

Figure Lengend Snippet: Figure 3. ER stress-induced apoptosis of ovarian cancer cells after downregulation of PHLDA1. Apoptosis assay of 2008 (A) and SKOV3 (B) cells expressing shctrl or shPHLDA1 after treatment with thapsigargin (Tg). **P<0.001 compared with the shctrl group. Mean ± SD, n = 3.

Techniques: Apoptosis Assay, Expressing

Journal: Journal of Cancer

Article Title: PHLDA1 Modulates the Endoplasmic Reticulum Stress Response and is required for Resistance to Oxidative Stress-induced Cell Death in Human Ovarian Cancer Cells.

doi: 10.7150/jca.45262

Figure Lengend Snippet: Figure 5. Expression of autophagy-related proteins, anti-apoptosis proteins, ER stress-associated proteins after PHLDA1-downregulation in ovarian cancer cells. (A) Western blot analysis of Autophagy-related proteins (Beclin-1, P62, and LC3) and anti-apoptosis protein (Bcl-2) in 2008 cells expressing shctrl or shPHLDA1 after treatment with 0.2 mM H2O2. (B and C) Western blot analysis of ER stress-associated proteins (IRE1α, PERK, BIP, ERO1-Lα, and PDI) in 2008 (B) and SKOV3 (C) cells after treatment with 0.2 mM H2O2.

Techniques: Expressing, Western Blot

Review

encoding scrambled shctrl (Addgene inc)

Structured Review

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1) Product Images from "PHLDA1 Modulates the Endoplasmic Reticulum Stress Response and is required for Resistance to Oxidative Stress-induced Cell Death in Human Ovarian Cancer Cells."

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Transduction:

Plasmid Preparation:

Transfection:

Construct:

Control:

shRNA:

Negative Control:

Expressing:

Cloning:

other:

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