Journal: Cell

Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

doi: 10.1016/j.cell.2023.12.026

Figure Lengend Snippet: (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial CaLu-3 cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Blots were then incubated with polyclonal anti-S2 antibody (Sino Biological, 40590; RRID:AB_2857932), anti-S1 antibody (Sino Biological, 40591-T62; RRID: AB_2893171), and anti-GAPDH as a loading control (Santa Cruz, Cat# sc-47724, RRID: AB_627678).

Techniques: Binding Assay, Membrane, Infection, Stable Transfection, Expressing, Derivative Assay, Transfection

Journal: Cell

Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

doi: 10.1016/j.cell.2023.12.026

Figure Lengend Snippet: (A and B) Cell surface expression of the indicated variant spike proteins. HEK293T cells used for production of pseudotyped lentiviral vectors carrying each variant spike proteins ( , , and ) were stained with anti-SARS-CoV-2 S1 antibody. Representative histogram of anti-S1 signals in the cells. (A) and geometric mean fluorescence intensities (B) of each subvariant from three biological replicates are shown. (C) Spike expression and processing in viral producer cell lysates. HEK293T cells, which were used to produce lentiviral pseudotypes, were lysed and probed with anti-S1, anti-S2, and anti-GAPDH antibodies, respectively. Spike processing was quantified by NIH ImageJ and set to a surface S1 (S1/S) or S2/S ratio and normalized the ratios of each Omicron subvariant to that of D614G. Dots represent three biological replicates. Bars in (B) represent means ± standard error. Significance relative to D614G was made using a one-way ANOVA with Bonferroni post-test. p values are displayed as ns p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: Blots were then incubated with polyclonal anti-S2 antibody (Sino Biological, 40590; RRID:AB_2857932), anti-S1 antibody (Sino Biological, 40591-T62; RRID: AB_2893171), and anti-GAPDH as a loading control (Santa Cruz, Cat# sc-47724, RRID: AB_627678).

Techniques: Expressing, Variant Assay, Staining, Fluorescence

Journal: Cell

Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

doi: 10.1016/j.cell.2023.12.026

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Blots were then incubated with polyclonal anti-S2 antibody (Sino Biological, 40590; RRID:AB_2857932), anti-S1 antibody (Sino Biological, 40591-T62; RRID: AB_2893171), and anti-GAPDH as a loading control (Santa Cruz, Cat# sc-47724, RRID: AB_627678).

Techniques: Infection, Recombinant, Transfection, Modification, Protease Inhibitor, Western Blot, Software, Imaging

Journal: bioRxiv

Article Title: Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants

doi: 10.1101/2021.04.01.437942

Figure Lengend Snippet: A, VH, VK and VL gene usage of antibodies from plasmablasts and memory B cells (MBCs). Up to the top four genes in each chart are shown with different colors (genes that were tied for 4 th and lower are not highlighted). B , Tukey’s plots showing heavy and light chain gene mutations of antibodies from plasmablasts and MBCs. Percent mutations were compared with the Mann-Whitney U-test. The middle line shows the median, and the box extends from the 1 st to 3 rd quartile. C , Top 21 neutralizing antibodies (IC 50 <1 μg/mL) by antigen specificity (left), and neutralization curves of selected antibodies (right). D , SARS-CoV-2 neutralization potency versus heavy chain mutation levels of antibody panel. The 5 most potent antibodies are shown as solid circles. E , Neutralization potency of antibodies by cell type. Top values indicate percentages of non-neutralizing antibodies. Horizontal bars indicate mean values; Mann-Whitney U-test (non-neutralizing antibodies excluded from calculation). The 5 most potent antibodies are shown in color. F , SARS-CoV-2 neutralization curves of benchmark antibodies from different groups , , . G, Neutralization IC 50 values of antibodies in our panel and benchmark antibodies in three different neutralization assays. Authentic SARS-CoV-2 FRNA values are from a single experiment done in quadruplicate, authentic SARS-CoV-2 (Scripps) values are an average of two experiments done in duplicate, pseudovirus (MLV) values are an average of 3 experiments in duplicate. The colors indicate the different sources of the antibodies: red, this study; blue, ref. ; yellow, ref. ; green, ref. .

Article Snippet: Following fixation, the cells were permeabilized with Triton X-100 and probed with a SARS-CoV/SARS-CoV-2 nucleoprotein-specific rabbit primary antibody (Sino Biological, Wayne, PA, USA, 40143-R001) followed by an Alexa Fluor 647-conjugated secondary antibody (Life Technologies, San Diego, CA, USA, A21245).

Techniques: MANN-WHITNEY, Neutralization, Mutagenesis

Journal: bioRxiv

Article Title: Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants

doi: 10.1101/2021.04.01.437942

Figure Lengend Snippet: A , Isoaffinity plot of antibodies targeting SARS-CoV-2 RBD (representative of n = 2 experiments). The affinity (K D ) values on the top and right of the plot refer to the dotted lines crossing the plot, e.g. any antibody falling on the 10 pM dotted line has a K D value of 10 pM. B , Neutralization potency versus affinity of anti-RBD antibodies. C , Isoaffinity plot of antibodies targeting SARS-CoV-2 NTD (representative of n = 2 experiments). The affinity (K D ) values on the top and right of the plot refer to the dotted lines crossing the plot, e.g. any antibody falling on the 10 pM dotted line has a K D value of 10 pM. D , Neutralization potency versus affinity of anti-NTD antibodies. E , Epitope binning of anti-RBD antibodies (representative of n = 2 experiments). ACE2 was only used as an analyte (competitor) and not as a ligand, while all other antibodies were tested as both ligands and analytes. Solid lines indicate two-way competition while dotted lines indicate one-way competition. The number and percentage of neutralizing antibodies (IC 50 < 10 μg/mL) in each bin are shown. F , Epitope bins represented by C135 (yellow bin), S309 (purple bin), ACE2 (red bin), CR3022 (cyan bin), as well as the NTD-specific antibody 4–8 (orange) modeled onto a SARS-CoV-2 spike protein (white cartoon). Antibody 4–8 was not binned successfully in our experiments but binds to a similar region to 2–17 and 5–24 (ref. ), which were binned. The epitope sites are color-coded the same as in Fig. 2E and 2G. N-glycans at the N343 glycan site are represented by sticks. G , Epitope binning of anti-NTD antibodies (representative of n = 2 experiments). All antibodies were tested as both ligands and analytes. Solid lines indicate two-way competition while dotted lines indicate one-way competition. The number and percentage of neutralizing antibodies (IC 50 < 10 μg/mL) in each bin are shown. H , Binding of mAb panel to spike protein containing mutations from B.1.1.7 and B.1.351 variants (n = 1 experiment). The numbers show the percentages of mAb binding to mutants relative to D614G (which was normalized to 100). I , Neutralization potency of CV503, CV664 and CV993 against B.1.1.7 and B.1.351 variants relative to wild-type (pseudotyped) SARS-CoV-2 (n = 1 experiment). Ratios are shown in brackets, and numbers smaller than 1 indicate an increase in potency while numbers larger than 1 indicate a decrease in potency relative to wild-type.

Article Snippet: Following fixation, the cells were permeabilized with Triton X-100 and probed with a SARS-CoV/SARS-CoV-2 nucleoprotein-specific rabbit primary antibody (Sino Biological, Wayne, PA, USA, 40143-R001) followed by an Alexa Fluor 647-conjugated secondary antibody (Life Technologies, San Diego, CA, USA, A21245).

Techniques: Neutralization, Binding Assay

Journal: bioRxiv

Article Title: Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants

doi: 10.1101/2021.04.01.437942

Figure Lengend Snippet: A , CV503 binds to the ridge region of SARS-CoV-2 RBD. The heavy and light chains of CV503 are shown in orange and yellow, respectively. SARS-CoV-2 RBD is in white, where its ridge region (residues 471–491) is shown in blue. B, The ACE2/RBD complex structure (PDB ID: 6M0J) is superimposed on the CV503/RBD complex. The heavy chain of CV503 (orange) would clash with ACE2 (green) if bound to RBD simultaneously (indicated by red circle). C-D , Epitope of CV503. Epitope residues contacting the heavy chain are in dark blue and light chain are in light blue, while residues contacting both heavy and light chains are in ocean blue. In C , CDR loops that are directly involved in RBD-binding are labeled. In D , epitope residues are labeled. Epitope residues that are also involved in ACE2 binding are labeled in red. E, ACE2-binding site on the RBD are in light pink. ACE2 is represented as semi-transparent cartoon in pale green. Epitope residues and ACE2-interacting residues are defined as those with a buried surface area (BSA) > 0 Å 2 . F , F486 at the ridge region of SARS-CoV-2 RBD (blue) is clamped in a hydrophobic pocket formed by the heavy (orange) and light chains (yellow) of CV503.

Article Snippet: Following fixation, the cells were permeabilized with Triton X-100 and probed with a SARS-CoV/SARS-CoV-2 nucleoprotein-specific rabbit primary antibody (Sino Biological, Wayne, PA, USA, 40143-R001) followed by an Alexa Fluor 647-conjugated secondary antibody (Life Technologies, San Diego, CA, USA, A21245).

Techniques: Binding Assay, Labeling

Journal: bioRxiv

Article Title: Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants

doi: 10.1101/2021.04.01.437942

Figure Lengend Snippet: A , Scheme of DVD-Ig ™ . In our bispecific antibody naming system, the first name refers to the antibody used to make the outer binding site and the second refers to the antibody at the inner binding site. GS or EL refers to the type of linker connecting the two antigen-binding sites. See for details. B , Binding of individual and bispecific antibodies to various domains from SARS-CoV-1 and SARS-CoV-2 (representative of n = 2 experiments). Area under the curve (AUC) values are shown after subtraction with the negative control antigen. C , Neutralization potency of bispecific antibodies with SARS-CoV-2 authentic and pseudotyped virus (MLV). Values are averaged from two experiments done in duplicate. D , Neutralization curves of CV1206_521_GS with SARS-CoV-2 authentic and pseudotyped virus. Curves are from a representative experiment, IC 50 values for authentic virus are the average from two experiments and those for the pseudovirus are from an average of two (bispecific) or three (regular antibody) experiments. E, Neutralization potency of CV1206_521_GS versus a cocktail of CV1206 and CV521, with concentrations shown in the molar scale to enable a fair comparison. For the antibody combination, the values on the x-axis refers to the concentration of each antibody in the cocktail, e.g. 10 nM refers to 10 nM of CV1206 + 10 nM of CV521. F , 3D reconstruction of CV1206_521_GS from nsEM images. Two ”one RBD up” models (PDB 6VYB) in green are docked into the reconstruction. Similarly, multiple mock scFv’s in orange and purple were docked to approximate the DVD-Ig molecule. O, outer binding site; I, inner binding site. G , Binding of bispecific antibody panel to spike protein containing mutations from B.1.1.7 and B.1.351 variants (n = 1 experiment). The numbers show the percentages of mAb binding to mutants relative to D614G (which was normalized to 100). H , Neutralization potency of bispecific antibodies against D614G, B.1.1.7 and B.1.351 variants relative to wild-type (pseudotyped) SARS-CoV-2 (n = 1 experiment). Ratios are shown in brackets: numbers smaller than 1 indicate an increase in potency while numbers larger than 1 indicate a decrease in potency relative to wild-type. ND, not determined. I, Potency of CV503_664_EL versus individual component mAbs against wild-type and B.1.351 SARS-CoV-2 pseudotyped virus (lentivirus).

Article Snippet: Following fixation, the cells were permeabilized with Triton X-100 and probed with a SARS-CoV/SARS-CoV-2 nucleoprotein-specific rabbit primary antibody (Sino Biological, Wayne, PA, USA, 40143-R001) followed by an Alexa Fluor 647-conjugated secondary antibody (Life Technologies, San Diego, CA, USA, A21245).

Techniques: Binding Assay, Negative Control, Neutralization, Concentration Assay

Journal: Biomedicines

Article Title: Morphological and Immunocytochemical Characterization of Paclitaxel-Induced Microcells in Sk-Mel-28 Melanoma Cells

doi: 10.3390/biomedicines12071576

Figure Lengend Snippet: Primary and secondary antibodies.

Article Snippet: ALDH2 , Primary antibody: rabbit anti-human Rabbit polyclonal , , 1:75 , 158-201549-T08-200, Sino Biological by Nordic BioSite, Täby, Sweden.

Techniques: Concentration Assay, Recombinant

Journal: Nature metabolism

Article Title: Glucose-dependent partitioning of arginine to the urea cycle protects β-cells from inflammation

doi: 10.1038/s42255-020-0199-4

Figure Lengend Snippet: a, Histogram of relative abundance of 3160 proteins detected (out of 5399 total, see ) by LC–MS/MS in 8.7 × 10 4 purified human ²-cells from n = 3 donors. Green lines indicate gene products related to the urea cycle, argininosuccinate and aspartate metabolism detected at the protein level. b, Expression levels of genes related to the urea cycle, pyruvate metabolism and arigninosuccinate shunt based on RNAseq analysis of FACS-sorted human β-cells from n=3 human donors, data are in Log2CPM. c, Co-immunostaining of insulin and individual metabolic enzymes related to the urea cycle in dispersed human islet cells from one donor representing similar results obtained from two additional donors. Representative images are shown (left), scale bar is 10 microns. Mean fluorescence intensity (FI) within the insulin positive region of interest (ROI) was calculated from 5 images per antibody (right). Neg denotes negative control for background Alexa Fluor 488 signal with insulin co-stain. Statistical analyses are student’s t-tests of each enzyme compared to Neg. Enzyme abbreviations in a–c are ARG2, arginase 2; ASL, argininosuccinate lyase; ASS1, argininosuccinate synthase 1; CPS1, carbamoyl-phosphate synthase 1; DDAH 1, dimethylarginine dimethylaminohydrolase 1; DDAH 2, dimethylarginine dimethylaminohydrolase 2; GOT1, aspartate aminotransferase 1; GOT2, aspartate aminotransferase 2; MDH1, malate dehydrogenase 1; MDH2, malate dehydrogenase 2; NAGS, N-acetyl-glutamate synthase; OAT, ornithine aminotransferase; PC, pyruvate carboxylase; SLC25A12, solute carrier family 25 member 12/calcium-binding mitochondrial carrier protein Aralar 1; SLC25A13, solute carrier family 25 member 13/calcium-binding mitochondrial carrier protein Aralar 2; SLC25A15, solute carrier family 25 member 15/mitochondrial ornithine transporter 1. d–e, Cytokine-induced changes in the partitioning of 15 N 2 -L-arginine to urea ( d ) and citrulline/NO ( e ) synthesis in human islets comparing protective vs non-protective glucose metabolism as modeled by BAD SAHB A SD vs RO0281675 treatment, respectively, n=4 donors. f, Chemical inhibition of arginase via ABH interferes with the protective effect of BAD SAHB A SD in human islets undergoing inflammation, n=5 donors. Data are means ± s.e.m. with one-way (d,e) and two-way (f) ANOVA statistical tests with Tukey adjustment for multiple comparisons.

Article Snippet: For all genetic manipulations, corresponding changes in protein expression were verified by western blot analysis using the following antibodies and dilutions; GOT1 (Sino Biological 14196-T52-50, 1:1000), GOT2 (My Bio Source MBS769801, 1:2000), glucokinase (Santa Cruz, 1:1000), pyruvate carboxylase (PCB, Santa Cruz, 1:1000), ARG2 (Life technologies, 1:1000), V5 (Cell signaling, 1:1000), MYC-tag (Cell Signaling, 1:1000), BAD (Abcam, 1:3000), and actin (Sigma, 1:20000).

Techniques: Liquid Chromatography with Mass Spectroscopy, Purification, Expressing, Immunostaining, Fluorescence, Negative Control, Staining, Binding Assay, Inhibition

Journal: Nature metabolism

Article Title: Glucose-dependent partitioning of arginine to the urea cycle protects β-cells from inflammation

doi: 10.1038/s42255-020-0199-4

Figure Lengend Snippet: a, Schematic of the TCA and urea cycles and their connection via the aspartate-argininosuccinate shunt. Enzymes of interest are marked in red and their corresponding inhibitors in blue. b, Total aspartate levels in human islets treated with the indicated compounds and cultured in the absence or presence of inflammatory cytokines. Data are from the untargeted metabolomics analysis in , n=5 human donors pooled and split into 8 replicates. c, Contribution of glucose to total aspartate pools in mouse islets labeled with 13 C 6 glucose and treated with inflammatory cytokines in the context of protective vs non-protective glucose metabolism. Data are from n=5 (Veh, RO0281675), n=6 (BAD SAHB A SD) and n=4 (BAD SAHB A AAA) independent mouse islet isolations and experiments. See for isotopologue distribution of aspartate in an analogous labelling experiment. d–e, Quantification of urea and NO , and viability ( e ) in human islets subjected to shRNA-mediated GOT1 ( G1 ) or GOT2 ( G2 ) depletion and treated with cytokines in the context of protective vs non-protective glucose metabolism, n=4 donors for urea, n=3 donors for NO, and n=5 donors for viability measurements. Data for one hairpin per gene are displayed (sh#1 for GOT1 and sh#2 for GOT2 ) from the full data set of multiple hairpins, see – . f–g, Quantification of urea levels ( f ) and viability ( g ) in human islets supplemented with argininosuccinate (AS) in the presence of inflammatory cytokines, H 2 O serves as vehicle control for AS. Data are from 6 independent experiments from n=3 donors. Data are means ± s.e.m. with statistical analyses on means from independent experiments using one-way ANOVA with Tukey adjustment for multiple comparisons.

Article Snippet: For all genetic manipulations, corresponding changes in protein expression were verified by western blot analysis using the following antibodies and dilutions; GOT1 (Sino Biological 14196-T52-50, 1:1000), GOT2 (My Bio Source MBS769801, 1:2000), glucokinase (Santa Cruz, 1:1000), pyruvate carboxylase (PCB, Santa Cruz, 1:1000), ARG2 (Life technologies, 1:1000), V5 (Cell signaling, 1:1000), MYC-tag (Cell Signaling, 1:1000), BAD (Abcam, 1:3000), and actin (Sigma, 1:20000).

Techniques: Cell Culture, Labeling, shRNA

Journal: Nature metabolism

Article Title: Glucose-dependent partitioning of arginine to the urea cycle protects β-cells from inflammation

doi: 10.1038/s42255-020-0199-4

Figure Lengend Snippet: a, 13 C fractional labelling of aspartate from 13 C 6 glucose. Data are shown as non-normalized to vehicle PBS and display the fraction of each M+ n mass isotopomer out of the total pool of aspartate for each condition. For clarity, statistical comparisons are only shown for each M+ n of a given condition (RO0281675, BAD SAHB A SD and BAD SAHB A AAA) compared to the corresponding M+ n of vehicle control. Data are pooled means from n=6 (Veh), n=5 (RO0281675), and n=6 (BAD SAHB A SD, BAD SAHB A AAA) independent mouse islet isolations and experiments. b, Western blot analysis of GOT1/2 knockdown efficiency using multiple independent hairpins for data shown in – and – . Blots are representative of n=2 independent experiments with similar results. c–d, Aspartate ( c ), urea and NO ( d ) levels in human islets from the same experiments shown in – , displaying the complete set of data on all hairpins tested. Aspartate data are from n = 4 human donors for shCtrl samples and n = 3 donors for knockdown samples. Urea and NO data are from n = 4 and n = 3 donors, respectively. Statistical analyses in (a) are two-way ANOVA showing p-value comparisons for each condition to Veh, and one-way ANOVA in (c–d), both with Tukey adjustment for multiple comparisons.

Article Snippet: For all genetic manipulations, corresponding changes in protein expression were verified by western blot analysis using the following antibodies and dilutions; GOT1 (Sino Biological 14196-T52-50, 1:1000), GOT2 (My Bio Source MBS769801, 1:2000), glucokinase (Santa Cruz, 1:1000), pyruvate carboxylase (PCB, Santa Cruz, 1:1000), ARG2 (Life technologies, 1:1000), V5 (Cell signaling, 1:1000), MYC-tag (Cell Signaling, 1:1000), BAD (Abcam, 1:3000), and actin (Sigma, 1:20000).

Techniques: Western Blot

Journal: Oncotarget

Article Title: The HGF inhibitory peptide HGP-1 displays promising in vitro and in vivo efficacy for targeted cancer therapy

doi:

Figure Lengend Snippet: a. K D of HGP-1 binding to HGF was determined by SPR technique. b. The assessment of binding competition between various proteins and HGF by fluorescence-based ELISA assay post 1.5-hour incubation. Proteins at the concentrations of 0.05 nM, 0.5 nM, 5 nM and 50 nM mixed with 10 μM FITC-labeled HGP-1 were the liquid phase ( n = 5). c. The binding activity between HGP-1 to HGF and EGF were measured by fluorescence-based direct ELISA assay post 1.5-hour incubation. HGP-1 at the concentrations of 0.1 μM, 1 μM, 10 μM, 100 μM were used ( n = 3). Values were mean ± SEM.

Article Snippet: Recombinant human full length HGF (10463- NHAC-A), VEGF-165 (11066-HNAB), Protein G (13103-PNAE), Biotinylated recombinant human full length HGF (10463-HNAC-B), HGF antibody (10463-RP01-B), MET antibody (10692-MM02), EGF antibody (0605-R008) were purchased from Sino Biological Inc. Recombinant human EGF (H6000–10), bFGF (H3000–10) were obtained from Beijing Wishbiotechnology Co., Ltd. Dynabeads MyOne streptavidin C1 magnetic beads (#65002) and Streptavidin Phycoerythrin Conjugates (S-866) were obtained from Invitrogen.

Techniques: Binding Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Incubation, Labeling, Activity Assay, Direct ELISA

Journal: Oncotarget

Article Title: The HGF inhibitory peptide HGP-1 displays promising in vitro and in vivo efficacy for targeted cancer therapy

doi:

Figure Lengend Snippet: The cells were treated with HGP-1 (5 μM and 50 μM) and SP-H1 (50 μM) for 15 minutes respectively. a. Representatives Western Blot of phospho-MET level in A549 cells after treated by HGF alone or combining with HGP-1. b. Quantitation of HGP-1 inhbitory effect on phospho-MET level in A549 cells. c. Representatives Western blot of phospho-ERK1/2 level in A549 cells after HGP-1 treatment. d. Quantitaion of HGP-1 inhibitory effect on phospho-ERK1/2 level in A549 cells. * P < 0.05, **0.01 < P < 0.05, represented the significance between HGF and HGP-1 or SP-H1 group. Values were mean ± SEM ( n = 5).

Article Snippet: Recombinant human full length HGF (10463- NHAC-A), VEGF-165 (11066-HNAB), Protein G (13103-PNAE), Biotinylated recombinant human full length HGF (10463-HNAC-B), HGF antibody (10463-RP01-B), MET antibody (10692-MM02), EGF antibody (0605-R008) were purchased from Sino Biological Inc. Recombinant human EGF (H6000–10), bFGF (H3000–10) were obtained from Beijing Wishbiotechnology Co., Ltd. Dynabeads MyOne streptavidin C1 magnetic beads (#65002) and Streptavidin Phycoerythrin Conjugates (S-866) were obtained from Invitrogen.

Techniques: Western Blot, Quantitation Assay