Journal: Clinical Cancer Research

Article Title: LAMC2 Regulates Key Transcriptional and Targetable Effectors to Support Pancreatic Cancer Growth

doi: 10.1158/1078-0432.ccr-22-0794

Figure Lengend Snippet: Figure 1. LAMC2 overexpression in human and mouse PDAC. A, Standardized mean difference (log2 scale) of the 8GS (LAMB3, DUSP6, AREG, LAMC2, FOSL1, PHLDA1, SPRY4, and DUSP4) mRNA expression across seven human PDAC datasets. B, Histoscore (H score) assessed for intensity and area expression of LAMC2 in a tissue arrayof 119 PDAC samples. C, Representative hematoxylin and eosin (H&E) and IHC for LAMC2 in human PDAC of different quartiles (Q4, quartile 4; Q3, quartile 3; Q2, quartile 2; Q1, quartile 1) and normal pancreas. Circled areas correspond to islets of Langerhans. Arrow points at a normal duct; scale bar, 100 mm. D, LAMC2 expression levels in multiple cell types extracted from a scRNA-seq analysis of human normal and PDAC samples. E, Western blot analysis of LAMC2 expression in a panel of 12 human PDAC cell lines. F, Representative IHC of LAMC2 (top), CK19 (center), and MUC5a (bottom) expression in normal pancreatic tissue (left), pancreatic intraepithelial neoplasia (PanIN; center), and PDAC (right); scale bar, 60 mm. G, LAMC2 immunoblot in a panel of 14 cell lines derived from KPf/fC and KPC mice.

Article Snippet: The following antibodies were used: LAMC2 (Santa Cruz Biotechnology, sc28330; RRID:AB_2134476), MUC5A (Thermo Fisher Scientific, MA5–12178; RRID:AB_10978001), CK19 (mouse: DSHB, TROMA III; RRID:AB_2133570), Ki67 (Cell Signaling Technology, 12202; RRID:AB_2620142), and FOSL1 (Cell Signaling Technology, 5281; RRID:AB_10557418).

Techniques: Over Expression, Expressing, Western Blot, Derivative Assay

Journal: Clinical Cancer Research

Article Title: LAMC2 Regulates Key Transcriptional and Targetable Effectors to Support Pancreatic Cancer Growth

doi: 10.1158/1078-0432.ccr-22-0794

Figure Lengend Snippet: Figure 2. LAMC2 inhibition impairs cell cycle and reduces cell viability. A, Western blot analysis to assess LAMC2 expression in PATU8902 and HPAFII cell lines in LAMC2 shRNA-mediated knocked-down and control cells. B, Cell proliferation assayed by MTS of the same cells as in A. C, Representative images (bottom) and quantification of colony forming ability in same cells as in A. D, LAMC2 immunoblot in 511950 and PDAC80 mouse cell lines in Lamc2 shRNA knocked-down or sgRNA knocked-out cells compared with control. E, Cell proliferation assayed by MTS of the same cells as in D. F, Representative images (bottom) and quantification of colony forming ability in the same cells as in D. G, Cell-cycle analysis by EdU labeling in the human cell lines after LAMC2 knockdown with a TET-inducible shRNA (1 mg/mL doxycycline). H, Apoptosis assessed by annexin V and 7AAD staining in both human cell lines upon LAMC2 knockdown. I, Cell-cycle analysis by EdU labeling in the mouse cell lines after Lamc2 shRNA-mediated knockdown. J, Apoptosis assessed by annexin V and 7AAD staining in mouse cell lines upon Lamc2 knockdown. Error bars correspond to SD in all experiments. All experiments were run at least three times.

Article Snippet: The following antibodies were used: LAMC2 (Santa Cruz Biotechnology, sc28330; RRID:AB_2134476), MUC5A (Thermo Fisher Scientific, MA5–12178; RRID:AB_10978001), CK19 (mouse: DSHB, TROMA III; RRID:AB_2133570), Ki67 (Cell Signaling Technology, 12202; RRID:AB_2620142), and FOSL1 (Cell Signaling Technology, 5281; RRID:AB_10557418).

Techniques: Inhibition, Western Blot, Expressing, shRNA, Control, Cell Cycle Assay, Labeling, Knockdown, Staining

Journal: Clinical Cancer Research

Article Title: LAMC2 Regulates Key Transcriptional and Targetable Effectors to Support Pancreatic Cancer Growth

doi: 10.1158/1078-0432.ccr-22-0794

Figure Lengend Snippet: Figure 3. Abrogation of LAMC2 invivo delaystumor growth.A and B, Immunoblot showingLAMC2 protein expression in a panelof 9 mouse PDAC celllines grown as 3D (A) and 2D (B) cultures. C and D, Organoid growth kinetics of 9 mouse cell lines grown in 3D (C) and 2D (D) assessed by CellTiter-Glo and MTS, respectively. Error bars correspond to SD. E and F, Representative images of organoids derived from mouse cell lines 511950 and PM12167 expressing control shGFP (top) and Lamc2 shRNA (bottom;E); scale bar, 100 mm. Growth kinetics assessed by CellTiter-Glo (F). Error bars correspond to SD. Invitro experimentswere done atleast three times. Gand H, Representative images of HPAFII cell line expressing a TET-inducible shRNA against LAMC2 (bottom) or a control shRNA (top) in 3D (G); scale bar, 100 mm. Growth kinetics assessed by CellTiter-Glo (H). Error bars correspond to SD. Invitro experimentswere done at least three times. I, Growth kinetics of subcutaneous (sc) tumors from PATU8902 (n ¼ 8) expressing a TET-inducible shRNA against LAMC2 or a control shRNA. Doxy: 2 mg/mL doxycycline in drinking water containing 5% sucrose. Error bars correspond to SEM. J, Dot plot showing tumor weight at sacrifice. Error bars correspond to SEM. (Continued on the following page.)

Article Snippet: The following antibodies were used: LAMC2 (Santa Cruz Biotechnology, sc28330; RRID:AB_2134476), MUC5A (Thermo Fisher Scientific, MA5–12178; RRID:AB_10978001), CK19 (mouse: DSHB, TROMA III; RRID:AB_2133570), Ki67 (Cell Signaling Technology, 12202; RRID:AB_2620142), and FOSL1 (Cell Signaling Technology, 5281; RRID:AB_10557418).

Techniques: Western Blot, Expressing, Derivative Assay, Control, shRNA

Journal: Clinical Cancer Research

Article Title: LAMC2 Regulates Key Transcriptional and Targetable Effectors to Support Pancreatic Cancer Growth

doi: 10.1158/1078-0432.ccr-22-0794

Figure Lengend Snippet: Figure 6. Inactivation of the LAMC2–AXL axis enhances MEK1/2i activity in 2D cultures. A, Cell proliferation assessed by MTS in PATU8902 and HPAFII human cell lines expressing a TET-inducible LAMC2 shRNA or shGFP control (1 mg/mL doxycycline) treated with or without the MEK1/2i trametinib at 5 nmol/L for 5 days. Error bars correspond to SD. B, Cell proliferation assessed by MTS in 511950 mouse cell lines expressing a Lamc2 or a control shRNA treated with or without the MEKi at 3 nmol/L (5111950) or 2 nmol/L (PM12167) for 5 days. Error bars correspond to SD. C, Cell proliferation in PATU8902 and HPAFII human cell lines treated with single MEKi at 2.5 nmol/L, AXLi (bemcentinib) at 0.5 mmol/L, or a combination of both, 5 days post-treatment. Error bars correspond to SD. D, Cell proliferation in 511950 and PDAC80 mouse cell lines treated with a MEKi (511950: 2 nmol/L; PDAC80: 1 nmol/L), an AXLi (0.2 mmol/L), or both, 5 days post-treatment. Error bars correspond to SD. E, Clonogenic assay images of PATU8902 and HPAFII cells treated with a MEKi (PATU8902: 1.25 nmol/L; HPAFII: 2.5 nmol/L), an AXLi (0.2 mmol/L), or both for 10 days. Average of experiments includes SD. F, Clonogenic assay images of mouse PDAC cell lines treated with a MEKi (511950: 1 nmol/L; PDAC80: 2.5 nmol/L), an AXLi (511950: 0.125 mmol/L; PDAC80: 0.25 mmol/L), or both for 10 days. Average of experiments includes SD. G and H. Western blot image of PARP in PATU8902 (G) and HPAFII (H) cells treated with MEKi (2.5 nmol/L), AXLi (0.5 mmol/L), combination of both, and vehicle control for 24 hours.

Article Snippet: The following antibodies were used: LAMC2 (Santa Cruz Biotechnology, sc28330; RRID:AB_2134476), MUC5A (Thermo Fisher Scientific, MA5–12178; RRID:AB_10978001), CK19 (mouse: DSHB, TROMA III; RRID:AB_2133570), Ki67 (Cell Signaling Technology, 12202; RRID:AB_2620142), and FOSL1 (Cell Signaling Technology, 5281; RRID:AB_10557418).

Techniques: Activity Assay, Expressing, shRNA, Control, Clonogenic Assay, Western Blot

Journal: Molecular Therapy Oncolytics

Article Title: The PABPC5/HCG15/ZNF331 Feedback Loop Regulates Vasculogenic Mimicry of Glioma via STAU1-Mediated mRNA Decay

doi: 10.1016/j.omto.2020.03.017

Figure Lengend Snippet: Endogenous Expression of PABPC5 and HCG15 and the Effect of PABPC5 and HCG15 on the Biological Behavior of Glioma Cells (A) PABPC5 protein expression levels in normal brain tissue (NBT), low-grade glioma tissue (LGGT), and high-grade glioma tissue (HGGT). The protein expression and corresponding IDV values of PABPC5 in NBT, LGGT, and HGGT are shown; the data are expressed as mean ± SD (n = 12). Compared with the NBT group, ∗∗p < 0.01; compared with the LGGT group, #p < 0.05. (B) PABPC5 protein levels in normal human astrocytes (NHAs) and U87 and U251 cells. The protein expression and corresponding IDV values of PABPC5 in NHAs and U87 and U251 cells are shown; the data are expressed as mean ± SD (n = 3). Compared with the NHA group, ∗p < 0.05. (C) The effect of knockdown of PABPC5 on the ability of proliferation was measured with CCK-8 in U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗p < 0.05. (D) The effect of knockdown of PABPC5 on the migration and invasion ability of U87 and U251 cells by was tested by Transwell test. Representative images and corresponding statistical analysis diagrams are shown. Data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗∗p < 0.01; the scale bar indicates 50 μm. (E) Three-dimensional culture was used to determine the effect of knockdown of PABPC5 on the VM formation ability of U87 and U251 cells. Representative images and corresponding statistical analysis plots are shown. Data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗∗p < 0.01. The scale bar indicates 50 μm. (F) Knockdown of PABPC5 regulates LAMC2 protein expression levels in U87 and U251 cells. The data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗p < 0.05 and ∗∗p < 0.01. (G) HCG15 expression in NBT, LGGT, and HGGT. Data are expressed as mean ± SD (n = 12). Compared with the NBT group, ∗∗p < 0.01 and ∗p < 0.05. Compared with the LGGT group, #p < 0.05. (H) HCG15 expression in NHAs and U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the NHA group, ∗∗p < 0.01 and ∗p < 0.05. (I) detecting the effects of knockdown and overexpression of HCG15 on proliferation. Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC group, ∗∗p < 0.01 and ∗p < 0.05. Compared with the HCG15 (+)-NC group, ##p < 0.01. (J) Detecting the effects of knockdown and overexpression of HCG15 on the migration and invasion ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–) -NC group, ∗∗p < 0.01. Compared with the HCG15 (+)-NC group, ## p < 0.01. The scale bar indicates 50 μm. (K) Effect of knockdown and expression of HCG15 on the VM formation ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC group, ∗∗p < 0.01. Compared with the HCG15 (+)-NC group, ##p < 0.01 and #p < 0.05. The scale bar indicates 50 μm. (L) Detecting the effects of knockdown and overexpression of HCG15 regulates LAMC2 protein expressional levels in U87 and U251 cells, respectively. Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC group, ∗∗p < 0.01. Compared with the HCG15 (+)-NC group, ##p < 0.01.

Article Snippet: The secondary antibodies were diluted (1:10,000, Proteintech, Rosemont, IL, USA); PABPC5 , ZNF331 , and LAMC2 were rabbit resistance, and GAPDH was mouse resistance.

Techniques: Expressing, Knockdown, CCK-8 Assay, Migration, Over Expression

Journal: Molecular Therapy Oncolytics

Article Title: The PABPC5/HCG15/ZNF331 Feedback Loop Regulates Vasculogenic Mimicry of Glioma via STAU1-Mediated mRNA Decay

doi: 10.1016/j.omto.2020.03.017

Figure Lengend Snippet: PABPC5 Enhanced the Biological Behavior of Glioma Cells by Increasing the Stability of HCG15 (A) Knockdown of PABPC5 affects HCG15 . Data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗∗p < 0.01. (B) Validating the binding interaction between PABPC5 and HCG15 . Data are expressed as mean ± SD (n = 3). Compared with the respective anti-normal IgG groups, ∗∗p < 0.01. (C) knockdown of PABPC5 to detect nascent HCG15 in U87 and U251 cells; data are expressed as mean ± SD (n = 3). (D) Effect of knockdown of PABPC5 on HCG15 half-life. Data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗∗p < 0.01 and ∗p < 0.05. (E) Detecting the effects of knockdown of PABPC5 , knockdown of HCG15 , and overexpression of HCG15 on the proliferation ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the control group, ∗∗p < 0.01. Compared with the PABPC5 (–) + HCG15 (–)-NC group, #p < 0.05 and ##p < 0.01. Compared with the PABPC5 (–) + HCG15 (+)-NC group, &&p < 0.05. (F) Detecting the effects of knockdown of PABPC5 , knockdown of HCG15 , and overexpression of HCG15 on the migration and invasion ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the control group, ∗p < 0.05 and ∗∗p < 0.01. Compared with the PABPC5 (–) + HCG15 (–)-NC group, # p < 0.05. Compared with the PABPC5 (–) + HCG15 (+)-NC group, &p < 0.05. The scale bar indicates 50 μm. (G) Detecting the effects of knockdown of PABPC5 , knockdown of HCG15 , and overexpression HCG15 on the VM ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the control group, ∗p < 0.05. Compared with the PABPC5 (–) + HCG15 (–)-NC group, #p < 0.05. Compared with the PABPC5 (–) + HCG15 (+)-NC group, &p < 0.05. The scale bar indicates 50 μm. (H) Detecting the effects of knockdown of PABPC5 , knockdown of HCG15 , and overexpression of HCG15 on LAMC2 protein expression in U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the control group, ∗p < 0.05 and ∗∗p < 0.01. Compared with the PABPC5 (–) + HCG15 (–)-NC group, #p < 0.05. Compared with the PABPC5 (–) + HCG15 (+)-NC group, &p < 0.05.

Article Snippet: The secondary antibodies were diluted (1:10,000, Proteintech, Rosemont, IL, USA); PABPC5 , ZNF331 , and LAMC2 were rabbit resistance, and GAPDH was mouse resistance.

Techniques: Knockdown, Binding Assay, Over Expression, Control, Migration, Expressing

Journal: Molecular Therapy Oncolytics

Article Title: The PABPC5/HCG15/ZNF331 Feedback Loop Regulates Vasculogenic Mimicry of Glioma via STAU1-Mediated mRNA Decay

doi: 10.1016/j.omto.2020.03.017

Figure Lengend Snippet: ZNF331 Was Lowly Expressed in Gliomas and Inhibited the Formation of Vascular Mimicry (A) ZNF331 protein expression levels in NBT, LGGT, and HGGT. Data are expressed as mean ± SD (n = 12). Compared with the NBT group, ∗∗p < 0.01. Compared with the LGGT group, ## p < 0.01. (B) ZNF331 protein expression levels in NHAs and U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the NHA group ∗∗p < 0.01. (C) Detecting the effects of knockdown and overexpression of ZNF331 on the proliferation ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗∗p < 0.01. Compared with the ZNF331 (+)-NC group, ##p < 0.01. (D) Detecting the effect of ZNF331 on the migration and invasion ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗p < 0.05 and ∗∗p < 0.01. Compared with the ZNF331 (+)-NC group, ##p < 0.01. The scale bar indicates 50 μm. (E) Detecting the effects of ZNF331 on the VM formation ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗∗p < 0.01. Compared with the ZNF331 (+)-NC group, #p < 0.05 and ##p < 0.01. The scale bar indicates 50 μm. (F) Detecting the effects of knockdown and overexpression of ZNF331 regulates LAMC2 protein expressional levels in U87 and U251 cells, respectively. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗∗p < 0.01 and ∗p < 0.05. Compared with the ZNF331 (+)-NC group, ##p < 0.01. (G) Expression levels of ZNF331 mRNA and protein are detected in U87 and U251 cells after knockdown of PABPC5 . Data are expressed as mean ± SD (n = 3). Compared with the PABPC5 (–)-NC group, ∗p < 0.05 and ∗∗p < 0.01. (H) The expression levels of ZNF331 mRNA and protein are detected in U87 and U251 cells after knockdown and overexpression of HCG15 . Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC group, ∗∗p < 0.01. Compared with the HCG15 (+)-NC group, #p < 0.05. (I) Expression levels of ZNF331 mRNA and protein in U87 and U251 cells after knockdown of PABPC5 , knockdown of HCG15 , and overexpression of HCG15 . Data are expressed as mean ± SD (n = 3). Compared with the control group, ∗p < 0.05 and ∗∗p < 0.01. Compared with the PABPC5 (–) + HCG15 (–)-NC group, #p < 0.05 and ##p < 0.01. Compared with the PABPC5 (–) + HCG15 (+)-NC group, &p < 0.05 and &&p < 0.01.

Article Snippet: The secondary antibodies were diluted (1:10,000, Proteintech, Rosemont, IL, USA); PABPC5 , ZNF331 , and LAMC2 were rabbit resistance, and GAPDH was mouse resistance.

Techniques: Expressing, Knockdown, Over Expression, Migration, Control

Journal: Molecular Therapy Oncolytics

Article Title: The PABPC5/HCG15/ZNF331 Feedback Loop Regulates Vasculogenic Mimicry of Glioma via STAU1-Mediated mRNA Decay

doi: 10.1016/j.omto.2020.03.017

Figure Lengend Snippet: HCG15 Degraded ZNF331 mRNA through the SMD Pathway, Enhancing the Biological Behavior of Glioma Cells (A) Predicted results of HCG15 binding site and dual-luciferase reporter gene assays in the 3′ UTR of ZNF331 mRNA. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331-3′UTR-Wt + HCG15 (+)-NC group, ∗p < 0.05. (B) RNA IP results. Data are expressed as mean ± SD (n = 3). Compared with the anti-normal IgG group, ∗∗p < 0.01. (C) The effect of HCG15 on the stability of ZNF331 mRNA; the data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC group, ∗∗p < 0.01. Compared with the HCG15 (+)-NC group, ##p < 0.01. (D) STAU1 effects on the stability of ZNF331 mRNA; data are expressed as mean ± SD (n = 3). Compared with the STAU1 (–)-NC group, ∗p < 0.05 and ∗∗p < 0.01. (E) The effect of UPF1 on the stability of ZNF331 mRNA; data are expressed as mean ± SD (n = 3). Compared with the UPF1 (–)-NC group, ∗p < 0.05 and ∗∗p < 0.01. (F) Effect of STAU1 on ZNF331 protein expression; data are expressed as mean ± SD (n = 3). Compared with the STAU1 (–)-NC group, ∗p < 0.05 and ∗∗p < 0.01. (G) Effect of UPF1 on ZNF331 protein expression; data are expressed as mean ± SD (n = 3). Compared with the UPF1 (–)-NC group, ∗p < 0.05. (H) Effect of HCG15 and STAU1 on ZNF331 protein expression; data are expressed as mean ± SD (n = 3). Compared with the control group, ∗p < 0.05 and ∗∗p < 0.01. Compared with the HCG15 (–) + STAU1 (–)-NC group, # p < 0.05 and ## p < 0.01. Compared with the HCG15 (–)+ STAU1 (+)-NC group, &p < 0.05 and &&p < 0.01. (I) Detecting the effects of HCG15 and ZNF331 on the proliferation ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC + ZNF331 (+)-NC group, ∗∗p < 0.01. (J) Examining the effects of HCG15 and ZNF331 on the migration and invasion ability of U87 and U251 cells. Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC + ZNF331 (+)-NC group, ∗∗p < 0.01 and ∗p < 0.05. The scale bar indicates 50 μm. (K) Detecting the effects of HCG15 and ZNF331 on the VM formation ability of U87 and U251 cells. Data are mean ± SD (n = 3). Compared with the HCG15 (–) + ZNF331 (+)-NC group, ∗∗p < 0.01. The scale bar indicates 50 μm. (L) Detecting the regulation of LAMC2 protein expression in U87 and U251 cells by HCG15 and ZNF331 . Data are expressed as mean ± SD (n = 3). Compared with the HCG15 (–)-NC + ZNF331 (+)-NC group, ∗∗p < 0.01.

Article Snippet: The secondary antibodies were diluted (1:10,000, Proteintech, Rosemont, IL, USA); PABPC5 , ZNF331 , and LAMC2 were rabbit resistance, and GAPDH was mouse resistance.

Techniques: Binding Assay, Luciferase, Expressing, Control, Migration

Journal: Molecular Therapy Oncolytics

Article Title: The PABPC5/HCG15/ZNF331 Feedback Loop Regulates Vasculogenic Mimicry of Glioma via STAU1-Mediated mRNA Decay

doi: 10.1016/j.omto.2020.03.017

Figure Lengend Snippet: ZNF331 Bound to the Promoter Regions of LAMC2 and PABPC5 (A) ZNF331 binds to the promoter of LAMC2 . Shown is a schematic diagram of the 2,000-bp LAMC2 promoter region upstream of the transcription start site (TSS), which is designated as +1. The putative ZNF331 binding site is shown. Using rabbit IgG as an NC, PCR was used to amplify the immunoprecipitated DNA. (B) ZNF331 binds to the promoter of PABPC5 . Shown is a schematic diagram of the 2,000-bp PABPC5 promoter region upstream of the TSS, which is designated as +1. The putative ZNF331 binding site is shown. Using rabbit IgG as an NC, PCR was used to amplify the immunoprecipitated DNA. (C) ZNF331 effect on LAMC2 mRNA expression. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗∗p < 0.01. Compared with the ZNF331 (+)-NC group, ##p < 0.01. (D) ZNF331 effects on expression of PABPC5 mRNA. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗∗p < 0.01. Compared with the ZNF331 (+)-NC group, ##p < 0.01. (E) ZNF331 affects the expression of PABPC5 protein. Data are expressed as mean ± SD (n = 3). Compared with the ZNF331 (–)-NC group, ∗∗p < 0.01 and ∗p < 0.05. Compared with the ZNF331 (+)-NC group, ## p < 0.01.

Article Snippet: The secondary antibodies were diluted (1:10,000, Proteintech, Rosemont, IL, USA); PABPC5 , ZNF331 , and LAMC2 were rabbit resistance, and GAPDH was mouse resistance.

Techniques: Binding Assay, Immunoprecipitation, Expressing

Journal: Cell

Article Title: Single-cell transcriptomic analysis of primary and metastatic tumor ecosystems in head and neck cancer

doi: 10.1016/j.cell.2017.10.044

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Monoclonal mouse LAMC2, clone CL2980, lot #CL2980 , Novus Biologicals , Cat#NBP2-42388.

Techniques: RNAscope, Labeling, Virus, Recombinant, Cloning, Transfection, Plasmid Preparation, Control, Software

Journal:

Article Title: A Mouse Model of Generalized non-Herlitz Junctional Epidermolysis Bullosa

doi: 10.1038/jid.2010.46

Figure Lengend Snippet: Expression of basement membrane zone proteins integrin alpha 6, collagen type XVII, and collagen type VII. Immunofluorescence of 129X1/SvJ +/+ and Lamc2jeb/Lamc2jeb tail skin using 63x lens on a Leica Confocal microscope at 546nm (red) adjusting threshold and gain to maximize dynamic range. Images on left are at 546nm on right are DAPI and bright field overlay. The blister cavity is indicated by a star. (bar = 50 μm) Dotted line represents dermal epidermal boundary.

Article Snippet: The following primary antibodies were used: rabbit anti mouse BP180 and COL7A1 (gifts of Z. Liu), rabbit anti mouse LAMA3, LAMB3, LAMC2 LE1-3 (AA 22-186, exons 2-6), LAMC2 LE4-6 (AA 460-606, exons 11-13, a gift of T. Sasaki) and rat anti-mouse ITGA6 (GeneTex, San Antonio, TX).

Techniques: Expressing, Membrane, Immunofluorescence, Microscopy