pkc θ  (Cell Signaling Technology Inc)

Journal: PLOS Pathogens

Article Title: Activating PKC-ε induces HIV expression with improved tolerability

doi: 10.1371/journal.ppat.1012874

Figure Lengend Snippet: Similar expression of PKC-θ in both T cells and platelets was observed, whereas PKC-η and ε were abundant in T cells but undetected in platelets. Cell lysates of isolated CD4 + T cells and platelets from healthy donors (n = 3) were tested for expression of novel PKC isoforms θ, η, and ε by western blotting utilizing PKC-θ and PKC-η/ε antibodies. Expression of GAPDH was used for normalization to enable accurate interpretation of differences in protein expression.

Article Snippet: For experiments shown in , primary antibodies against PKC-ɛ and -ɳ (LSBio, catalog C172661, 1:2000 dilution), PKC-θ (Cell Signaling Technology, catalog 13643, 1:1000 dilution), and GAPDH (Cell Signaling Technology, catalog 5174S, 1:1000 dilution) at RT for 60 minutes and secondary antibodies rabbit anti-mouse IgG-HRP (Santa Cruz Biotech, catalog sc-358914, 1:2000) and anti-rabbit IgG-HRP (Cell Signaling Technology, catalog 7074, 1:2000 dilution) at RT for 60 minutes were used.

Techniques: Expressing, Isolation, Western Blot

Journal: PLOS Pathogens

Article Title: Activating PKC-ε induces HIV expression with improved tolerability

doi: 10.1371/journal.ppat.1012874

Figure Lengend Snippet: Selectivity of PKC inhibitors towards PKC isoforms a .

Article Snippet: To study reactivation of latent cells by various PKC isoforms, 30 μg of pcDNA3.1 RFP vectors containing either PKC-ε, -η, -θ, or -δ, were transfected (GenScript’s Neon transfection system) according to the manufacturer’s instructions [ ].

Techniques:

Journal: PLOS Pathogens

Article Title: Activating PKC-ε induces HIV expression with improved tolerability

doi: 10.1371/journal.ppat.1012874

Figure Lengend Snippet: C-232A exhibits selectivity for novel PKC isoforms.

Article Snippet: To study reactivation of latent cells by various PKC isoforms, 30 μg of pcDNA3.1 RFP vectors containing either PKC-ε, -η, -θ, or -δ, were transfected (GenScript’s Neon transfection system) according to the manufacturer’s instructions [ ].

Techniques: Translocation Assay

Journal: PLOS Pathogens

Article Title: Activating PKC-ε induces HIV expression with improved tolerability

doi: 10.1371/journal.ppat.1012874

Figure Lengend Snippet: C-233 exhibits PKC-ε selectivity.

Article Snippet: To study reactivation of latent cells by various PKC isoforms, 30 μg of pcDNA3.1 RFP vectors containing either PKC-ε, -η, -θ, or -δ, were transfected (GenScript’s Neon transfection system) according to the manufacturer’s instructions [ ].

Techniques:

Journal: Cell Reports Methods

Article Title: A thermoplastic chip for 2D and 3D correlative assays combining screening and high-resolution imaging of immune cell responses

doi: 10.1016/j.crmeth.2025.100965

Figure Lengend Snippet:

Article Snippet: Rabbit anti-human PKC-Θ , Santa Cruz Biotechnology , Cat#sc-212; polyclonal C-18.

Techniques: Virus, Recombinant, Transfection, Cell Isolation, Software

Journal: Science Advances

Article Title: Molecular dynamics at immune synapse lipid rafts influence the cytolytic behavior of CAR T cells

doi: 10.1126/sciadv.adq8114

Figure Lengend Snippet: ( A ) Z -projection confocal microscopy of CD19-CAR T cells conjugated with Daoy.CD19 (20 min), CAR [emerald GFP (green)], and PKCθ (red) and transmitted light (TL). ( B and C ) Quantitation of (B) PKCθ and (C) CAR accumulated at CARIS. n = 48. ( D and E ) WB for total PKCθ in CD19-CAR T cells at baseline and after 20 min of recombinant CD19 stimulation (D), quantified in (E). ( F ) Schematic representation for lytic granule convergence to the MTOC and MTOC polarization to CARIS. Created using Biorender.com. ( G ) Time-lapse imaging of CD19-CAR T cells seeded on CD19 Fc + anti-CD18–coated glass, probed for lytic granules (LysoTracker) and MTOC (SiR-tubulin). ( H ) Change in the mean distance between lytic granules and the MTOC per cell of CD19-CAR CD28ζ (red line) and CD19-CAR 4-1BBζ (blue line) T cells. n = 32. ( I ) ImageStream for HER2-CAR T cells + LN229 (30 min). CARs [emerald GFP (green)], actin [phalloidin (red)], and MTOC [pericentrin (blue)]. ( J ) Distance between MTOC and CARIS. n = 360. Dot, conjugate [(B), (C), and (J)]. ( K and L ) Time-lapse tracking of distance of lytic granule centroid (LysoTracker) and MTOC (SiR-tubulin) from CARIS of CD19-CAR CD28ζ T cells + Daoy.CD19 (K) and SYTOX (target cell death); quantification of the percentage of CAR.emeraldGFP at CARIS (green), MTOC distance to CARIS (turquoise), and lytic granule centroid distance to CARIS (red) (L). Mean + SEM. n = 15. ( M and N ) CD107a flow cytometry on HER2-CAR (M) and CD19-CAR (N) T cells cocultured with LN229 and Daoy.CD19, respectively. ( O and P ) Supernatant of HER2-CAR T cells + LN299 (O) and CD19-CAR T cells + Daoy.CD19 (P) tested for perforin using ELISA. ( Q and R ) FasL flow cytometry on HER2-CAR (Q) and CD19-CAR (R) T cells cocultured with LN229 and Daoy.CD19, respectively. Student’s t test [(B) and (C)], RM two-way ANOVA [(H), (M), (N), (Q), and (R)], or one-way ANOVA [(J), (O) and (P)], with Holm-Sidak corrected multiple comparisons. n.s. = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The reagents/antibodies used were as follows: anti-human LAT (CST), AF647 CTB (Invitrogen, Carlsbad, CA), AF568-conjugated phalloidin (Life Technologies, Carlsbad, CA), anti-pericentrin rabbit polyclonal antibody (Abcam), AF647-conjugated mouse anti-perforin clone δG9 (BD), and anti-PKCθ antibody (Santa Cruz Biotechnologies, Dallas, TX).

Techniques: Confocal Microscopy, Quantitation Assay, Recombinant, Imaging, Flow Cytometry, Enzyme-linked Immunosorbent Assay