phosphate buffered saline pbs solution  (Bio-Rad)

Journal: The AAPS Journal

Article Title: Pharmacokinetics, Lymph Node Uptake, and Mechanistic PK Model of Near-Infrared Dye-Labeled Bevacizumab After IV and SC Administration in Mice

doi: 10.1208/s12248-012-9342-9

Figure Lengend Snippet: a SDS-PAGE of blank LN homogenate, free dye, bevacizumab in PBS (2 μg), bevacizumab–800CW conjugate (2 μg) in PBS and bevacizumab–800CW conjugate (2 μg) spiked in blank mouse lymph node

Article Snippet: Briefly, 8 μL of 500 μg/mL of bevacizumab IRDye 800CW conjugate or 500 μg/mL of bevacizumab in PBS solution was mixed with 12 μL of sample buffer (Bio-Rad, 2% SDS w/v , 25% glycerol v/v , 0.001% bromophenol blue w/v , 62.5 mM Tris–HCl, pH 6.8) with 5% β-mercaptoethanol, and 10 μL of each mixture was loaded and separated on a 10% polyacrylamide gel.

Techniques: SDS Page

Journal: Hepatology Communications

Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

doi: 10.1002/hep4.1181

Figure Lengend Snippet: Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).

Article Snippet: At different time points, culture media was removed and alamarBlue (BUF012A; Bio‐Rad, Hercules, CA) solution (1:10 dilution in phosphate‐buffered saline) was added to the cells.

Techniques: Western Blot, Infection, shRNA, Alamar Blue Assay, Fluorescence

Journal: Hepatology Communications

Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

doi: 10.1002/hep4.1181

Figure Lengend Snippet: Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).

Article Snippet: At different time points, culture media was removed and alamarBlue (BUF012A; Bio‐Rad, Hercules, CA) solution (1:10 dilution in phosphate‐buffered saline) was added to the cells.

Techniques: Western Blot, Alamar Blue Assay, Infection, shRNA, Cell Culture, Staining, Fluorescence

Journal: Hepatology Communications

Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

doi: 10.1002/hep4.1181

Figure Lengend Snippet: Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).

Article Snippet: At different time points, culture media was removed and alamarBlue (BUF012A; Bio‐Rad, Hercules, CA) solution (1:10 dilution in phosphate‐buffered saline) was added to the cells.

Techniques: Western Blot, Infection, shRNA, Alamar Blue Assay, Fluorescence

Journal: Hepatology Communications

Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

doi: 10.1002/hep4.1181

Figure Lengend Snippet: Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).

Article Snippet: At different time points, culture media was removed and alamarBlue (BUF012A; Bio‐Rad, Hercules, CA) solution (1:10 dilution in phosphate‐buffered saline) was added to the cells.

Techniques: Western Blot, Alamar Blue Assay, Infection, shRNA, Cell Culture, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Low doses of bioherbicide favour prion aggregation and propagation in vivo

doi: 10.1038/s41598-018-25966-9

Figure Lengend Snippet: Mice treated with 5 mg/kg of A6 compound exhibit a decreased lifespan and an increased PrP Sc burden. ( a ) Table summarizing the different groups of animals: prion inoculations, treatment performed, number of sick animals out of total number of animals (*means that in this group, one mouse was sacrificed while asymptomatic) and median survival time ± IQR (interquartile range). (b) Kaplan-Meier survival curves of mice intra-cerebrally (i.c.) inoculated with 5 μL of 22L prions, and treated with 50 μL of DMSO (n = 9, blue) or with 5 mg/kg of A6 (n = 9, red) by intraperitoneal (i.p.) route, (+22L + DM versus +22L + A6–5: non–parametric Mantel-Cox log-rank test, ** p- value = 0,0067). Healthy control mice, inoculated with 5 μL of PBS, were treated with 5 mg/kg of A6 (n = 5, black dashed, −22L + A6–5) or non-treated (n = 5, brown, −22L + DM). (c) Left panel: PET-blots analysis of coronal sections of control healthy mice non-inoculated with prions and treated with 5 mg/kg of A6 (−22L + A6–5) (1) ; mice inoculated with 22L prions and either treated with DMSO (+22L + DM) (2) or treated with A6 (+22L + A6–5) (3) . Mice were all sacrificed at the same time (149, 149 and 147 dpi respectively) either in an asymptomatic or sick stage. Right panel: PET-blots analysis of coronal sections of sick mice: +22L + DM (4) and +22L + A6–5 (5) sacrificed at the same time (162 and 163 dpi, respectively). Sections were labelled with SAF84 antibody to detect PrP Sc and revealed with Vectastain ABC-AmP kit.

Article Snippet: Brains from a terminally sick mouse (infected with 22L prions) or from a non-infected mouse were homogenized in 10% (w/v) sterile PBS solution using microbead-containing tubes and a Ribolysor apparatus (Biorad, Marnes la Coquette, France).

Techniques: Mouse Assay, Positron Emission Tomography