FIGURE 2.

Global kinome analysis of THP-1 cells infected with M. tuberculosis and ΔptpA M. tuberculosis. Simultaneous detection of selected host proteins and their activation status using a multiple immunoblotting technique are shown. Accurate intensity values for each protein are the accumulated signal obtained over a given scan time for each blot. These are shown as numerical values in Table 5. The three gels represent uninfected THP-1 cells (A), THP-1 cells infected with M. tuberculosis (B), and THP-1 cells infected with ΔptpA M. tuberculosis (C). Each lane was probed with one or more antibodies. The highlighted proteins are host signaling proteins showing a phosphorylation change greater than 25%. Antibodies against the phosphorylated proteins were as follows: lane 1, molecular size standard; lane 2, NR1 (Ser⁸⁹⁶); lane 3, PKR1 (Thr⁴⁵¹); lane 4, STAT5A (Tyr⁶⁹⁴); lane 5, PKCα (Ser⁶⁵⁷) and SRC (Tyr⁴¹⁸); lane 6, JNK (Thr¹⁸³ + Tyr¹⁸⁵) and RSK1/3 (Thr³⁵⁹ + Ser³⁶³/Thr³⁵⁶ + Ser³⁶⁰); lane 7, MEK3/6 (Ser¹⁸⁹/Ser²⁰⁷) and PKCα/β2 (Thr⁶³⁸ + Thr⁶⁴¹); lane 8, ERK1 (Thr²⁰² + Tyr²⁰⁴), ERK2 (Thr¹⁸⁵ + Tyr¹⁸⁷), S6Kα P70 (Thr³⁸⁹), and S6Kα P85 (Thr³⁸⁹); lane 9, PKCϵ (Ser⁷²⁹) and SMAD1/5/9 (Ser⁴⁶³ + Ser³⁶³/Ser⁴⁶³ + Ser⁴⁶⁵/Ser⁴⁶⁵ + Ser⁴⁶⁷); lane 10, STAT3 (Ser⁷²⁷); lane 11, JUN (Ser⁷³); lane 12, RAF1 (Ser²⁵⁹), STAT1α (Tyr⁷⁰¹), and STAT1β (Tyr⁷⁰¹); lane 13, PKBα (Akt1) (Thr³⁰⁸) and PKCδ (Thr⁵⁰⁷); lane 14, PKBα (Akt1) (Ser⁴⁷³); lane 15, GSK3α (Ser²¹), GSK3β (Ser⁹), and MSK1 (Ser³⁷⁶); lane 16, adducin α (Ser⁷²⁶), adducin γ (Ser⁶⁹³), CDK1/2 (Tyr¹⁵), and SRC (Tyr⁵²⁹); lane 17, GSK3α (Tyr²⁷⁹) and GSK3β (Tyr²¹⁶); lane 18, p38α MAPK (Thr¹⁸⁰ + Tyr¹⁸²) and RB (Ser⁷⁸⁰); lane 19, NPM (Ser⁴), and MEK1/2 (Ser²¹⁸ + Ser²²²); and lane 20, CREB1 (Ser¹³³) and RB (Ser⁸⁰⁷ + Ser⁸¹¹).